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WO2025012428A1 - Cartouche de manipulation d'au moins une substance, articles et procédés correspondants - Google Patents

Cartouche de manipulation d'au moins une substance, articles et procédés correspondants Download PDF

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Publication number
WO2025012428A1
WO2025012428A1 PCT/EP2024/069812 EP2024069812W WO2025012428A1 WO 2025012428 A1 WO2025012428 A1 WO 2025012428A1 EP 2024069812 W EP2024069812 W EP 2024069812W WO 2025012428 A1 WO2025012428 A1 WO 2025012428A1
Authority
WO
WIPO (PCT)
Prior art keywords
fluidic
channel
cartridge
unit
movable
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2024/069812
Other languages
English (en)
Inventor
Gerd Lüdke
Hassan Motejadded
Roman THIEL
Andreas Boos
Wolfgang Maurer
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Curetis GmbH
Original Assignee
Curetis GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Curetis GmbH filed Critical Curetis GmbH
Publication of WO2025012428A1 publication Critical patent/WO2025012428A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502738—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by integrated valves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0621—Control of the sequence of chambers filled or emptied
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/088—Channel loops
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0622—Valves, specific forms thereof distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0633—Valves, specific forms thereof with moving parts
    • B01L2400/0644—Valves, specific forms thereof with moving parts rotary valves

Definitions

  • the present invention relates to a cartridge for handling at least one substance, e.g. a biological substance such as microbiological substance. Moreover, the invention relates to corresponding subassemblies of the cartridge, e.g. to a stationary unit and to a movable unit. Furthermore, the invention relates to a method for preparing at least one molecular biological test such as microbiological test or biochemical test and to a method for performing at least one molecular biological test such as microbiological test or biochemical test, preferably using the cartridge.
  • micro-reactors The technical field of micro-reactors is a fast-growing technical field having applications not only in detection of human microbial infection or human virus diseases but also in agriculture, environmental protection, forensic, etc.
  • many micro-reactors have disadvantages, e.g. too cost expensive, too big, involving too much operator effort, impractical, etc.
  • Usage of cartridges may ease the usage of a machine in order to perform a plurality of tests.
  • the cartridges may prevent cross contamination of test substances used to perform the test.
  • the cartridge may provide the fluidic system of the micro-reactor, whereas the machine/device may provide driving units, heating units, stirring units etc.
  • the cartridge may be arranged within the device, e.g. within a cartridge receiving or cartridge retaining space.
  • a cartridge for handling at least one substance comprising:
  • a stationary unit comprising a stationary fluidic system
  • a movable unit comprising a movable fluidic system, wherein the movable unit is movable relative to the stationary unit, wherein the stationary fluidic system and the movable fluidic system are provided to receive, to guide, and/or to retain a fluid, wherein at least one of the stationary fluidic system and the movable fluidic system comprises a coupling portion, wherein the coupling portion comprises a fluidic passage, an intermediate volume and at least two fluidic interface portions, wherein in at least two different relative positions of the movable unit relative to the stationary unit, the cartridge is configured to establish a fluidic communication between the fluidic passage and a lumen, preferably a channel, of the fluidic system to which the coupling portion does not belong via a fluidic flow passing first through the fluidic passage, then through the intermediate volume and thereafter through a respective one of the at least two fluidic interface portions or vice versa.
  • the terms used herein are defined as described in “A multilingual glossary of biotechnological terms: (TUPAC Recommendations)”, Leuenberger, H.G.W, Nagel, B. and Kolbl, H. eds. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland).
  • the term “about” indicates a certain variation from the quantitative value it precedes.
  • the term “about” allows a ⁇ 5% variation from the quantitative value it precedes, unless otherwise indicated or inferred.
  • the use of the term “about” also includes the specific quantitative value itself, unless explicitly stated otherwise. For example, the expression “about 80°C” allows a variation of ⁇ 4°C, thus referring to range from 76°C to 84°C.
  • the present invention relates to a cartridge for handling at least one substance.
  • the cartridge provides a simple and uncomplex design, and in particular a design which can be inexpensively produced.
  • the invention also relates to a cartridge which can be used as a "disposable", i.e. a lab on a chip which is disposed after use.
  • the cartridge of the invention is particularly suitable for laboratory, in-field and point-of-care settings, e.g. for point-of-care testing (POCT).
  • POCT point-of-care testing
  • a laboratory may allow to provide and to monitor and/or to document usage of higher quality standards, e.g. for regulatory purposes.
  • the integration of all elements which will contact the at least one substance during handling in a - preferably disposable - unit further allows for the creation of a closed fluidic system, which helps preventing any contamination of the substances or of the interior of the device itself. Especially, cross contaminations may be prevented between different cartridges, especially between different cartridges or between substances and/or chemicals of different cartridges.
  • the chamber of the cartridge can be pre-filled with reagents adapted to perform a distinct handling. Therewith the cartridge can be used in a "ready-to-use" format of a lab on a chip.
  • POCT point-of-care testing
  • a biological testing such as medical diagnostic testing, or chemical testing at or near the point of care that is the time and place of individuals care, specifically patients care. This contrasts with the historical pattern in which testing was wholly or mostly confined to the medical or chemical laboratory, which entailed sending off specimens away from the point of care and then waiting hours or days to learn the results, during which time care must continue without the desired information.
  • Point-of-care tests are simple biological tests, such as medical tests, or chemical tests that can be performed at or near the bedside. The driving notion behind POCT is to bring the test conveniently and immediately to the individual, specifically patient.
  • POCT is often accomplished through the use of transportable, portable, and handheld instruments and test kits. Small bench analyzers or fixed equipment can also be used when a handheld device is not available - the goal is to collect the specimen and obtain the results in a very short period of time at or near the location of the individual, specifically patient, so that the treatment plan can be adjusted as necessary before the individual, specifically patient, leaves.
  • automation of the test results in many advantage, e.g. reduced costs, reduced time, reduced stuff, etc.
  • advantages of automation may be used in the laboratory as well as on a POCT.
  • integrated refers to a device that integrates one or several laboratory functions on a single carrier, e.g. on at least one plate of the cartridge.
  • Volumes that may be used in molecular biological tests may be in the range of 100 milliliter to 400 milliliter or in the range of 150 milliliter to 300 milliliter, to give only some examples. These volumes may be appropriate to provide enough volume of the substance, that may be only sparsely be present in a sample, e.g. in infectiology. However, if the concentration of the substance (under test) is high enough, the degree of integration may be raised.
  • LOC refers to a device that integrates one or several laboratory functions on a single integrated circuit (commonly called a "chip") of only millimeters to a few square centimeters, e.g. less than 20 cm 2 , to achieve automation and high-throughput screening or testing.
  • LOCs can handle extremely small fluid volumes down to less than 10 milliliter, less than 1 milliliter or less than 100 nano-liters. LOCs may use micro fluidics, the physics, manipulation and study of minute amounts of fluids. However, strictly regarded "lab-on-a-chip" indicates generally the scaling of single or multiple lab processes down to chip-format. Lower volumes may be applied for other medical/technical fields than infectiology or if the sample is concentrated in a preparation method.
  • the at least one substance may be a biological substance, such as a microbiological substance.
  • the term “at least one substance”, according to the present invention, refers to a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • a substance specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule) (from a microbial species), which is associated with a particular disease or condition or with a specific disease or condition stage.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • microbial species from a microbial species
  • the at least one substance e.g. nucleic acid molecule such as DNA or RNA molecule
  • a radiation transfer unit RU
  • a detectable dye specifically fluorescence marker/probe such as fluorophore
  • the nucleic acid molecule such as DNA or RNA molecule as the at least one substance is labelled with a detectable dye specifically fluorescence marker/probe such as TaqMan probe.
  • a TaqMan probe is a hydrolysis probe that is designed to increase the specificity of quantitative PCR.
  • the at least one substance is part of a sample such as biological sample as described herein. If the sample contains cellular material and the at least one substance (e.g. nucleic acid molecule such as DNA or RNA molecule) is contained therein, the cellular material needs to be lysed first in order to release the at least one substance (e.g. nucleic acid molecule such as DNA or RNA molecule) from the cells. Subsequently, the at least one substance (e.g. nucleic acid molecule such as DNA or RNA molecule) is isolated from the cell debris and then purified. In case the sample already contains the at least one substance (e.g.
  • nucleic acid molecule such as DNA or RNA molecule
  • a PCR reaction is preformed to amplify the nucleic acid molecule such as DNA or RNA molecule before detection.
  • the PCR reaction is preferably conducted in the presence of a TaqMan probe.
  • the RNA is first transcribed into cDNA before the amplification reaction is performed.
  • a TaqMan probe consists of a fluorophore covalently attached to the 5’-end of the oligonucleotide probe and a quencher at the 3 ’-end.
  • fluorophores e.g. 6- carboxyfluorescein, acronym: FAM, or tetrachlorofluorescein, acronym: TET
  • quenchers e.g. tetramethylrhodamine, acronym: TAMRA
  • the quencher molecule quenches the fluorescence emitted by the fluorophore when excited by the cycler’s light source via Forster resonance energy transfer (FRET).
  • TaqMan probes are designed such that they anneal within a nucleic acid such as DNA region amplified by a specific set of primers.
  • TaqMan probes can be conjugated to a minor groove binder (MGB) moiety, dihydrocyclopyrroloindole tripeptide (DPI3), in order to increase its binding affinity to the target sequence; MGB-conjugated probes have a higher melting temperature (T m ) due to increased stabilization of van der Waals forces.
  • MGB minor groove binder
  • DPI3 dihydrocyclopyrroloindole tripeptide
  • the 5' to 3' exonuclease activity of the Taq polymerase degrades the probe that has annealed to the template. Degradation of the probe releases the fluorophore from it and breaks the proximity to the quencher, thus, relieving the quenching effect and allowing fluorescence of the fluorophore.
  • fluorescence detected in the quantitative PCR thermal cycler is directly proportional to the fluorophore released and the amount of nucleic acid such as DNA template present in the PCR. This signal can then be detected with the radiation transfer unit (RU) mentioned above.
  • disease refers to an abnormal condition that affects the body of an individual.
  • a disease is often construed as a medical condition associated with specific symptoms and signs.
  • the term “disease” is often used more broadly to refer to any condition that causes pain, dysfunction, distress, social problems, or death to the individual afflicted, or similar problems for those in contact with the individual. In this broader sense, it sometimes includes injuries, disabilities, disorders, syndromes, infections, deviant behaviors, and atypical variations of structure and function, while in other contexts and for other purposes these may be considered distinguishable categories. Diseases usually affect individuals not only physically, but also emotionally, as contracting and living with many diseases can alter one’s perspective on life, and one’s personality.
  • the at least one substance is preferably associated with infectious diseases, inflammatory diseases, sepsis, autoimmune diseases, cancer diseases (or simply cancer), or any combinations thereof.
  • infectious disease refers to any disease which can be transmitted from individual to individual or from organism to organism, and is caused by a microbial agent (e.g. common cold). Infectious diseases are known in the art and include, for example, a viral disease, a bacterial disease, or a parasitic disease. Said diseases are caused by a virus, a bacterium, a fungus, and/or a parasite. The substance that causes an infectious disease can also be designated as pathogen.
  • pathogen refers to a virus, a bacterium, a fungus, and/or a parasite that can cause an infectious disease. It is then a pathogenic agent or substance.
  • the infectious disease is a respiratory disease such as pneumonia like hospitalized pneumonia, an implant or tissue infection, an intra-abdominal infection, or a urinary tract infection or a blood stream infection (e.g. if no other source of infection may be localized) or a CNS infection (meningitis/encephalitis).
  • a respiratory disease such as pneumonia like hospitalized pneumonia, an implant or tissue infection, an intra-abdominal infection, or a urinary tract infection or a blood stream infection (e.g. if no other source of infection may be localized) or a CNS infection (meningitis/encephalitis).
  • respiratory diseases refers to any disease affecting the respiratory system.
  • respiratory diseases include (i) obstructive lung diseases, (ii) restrictive lung diseases, (iii) respiratory tract infections, such as upper respiratory tract infections, e.g., common cold, sinusitis, tonsillitis, otitis media, pharyngitis, or laryngitis, and lower respiratory tract infections, e.g., pneumonia, (iv) respiratory tumors, e.g., small cell lung cancer, non-small cell lung cancer (e.g., adenocarcinoma, large cell undifferentiated carcinoma), other lung cancers such as carcinoid, Kaposi’s sarcoma, or melanoma, lymphoma, head and neck cancer, mesothelioma, and cancer metastasis in the lung such as from breast cancer, colon cancer, prostate cancer, germ cell cancer, and renal cell carcinoma, (v) pleural cavity diseases,
  • respiratory diseases that can be diagnosed using molecular diagnostics, preferably using nucleic acid amplification and analysis methods.
  • respiratory tract infections such as infections with pathogens, e.g., bacteria, viruses, yeast, or fungi, preferably yeast or bacteria, and respiratory tumors are preferred respiratory diseases in the context of the present invention.
  • Particularly more preferred respiratory diseases in the context of the present invention are pneumonias, in particular pneumonias caused by infections with pathogens, such as bacterial, viral, fungal, parasitic, atypical, community-acquired, healthcare-associated, hospital-acquired, ventilator-acquired pneumonia, or severe acute respiratory syndrome, tuberculosis, bronchitis, pathogenic infections during cystic fibrosis or chronic obstructive pulmonary disease (COPD), and a respiratory tumor.
  • pathogens such as bacterial, viral, fungal, parasitic, atypical, community-acquired, healthcare-associated, hospital-acquired, ventilator-acquired pneumonia, or severe acute respiratory syndrome, tuberculosis, bronchitis, pathogenic infections during cystic fibrosis or chronic obstructive pulmonary disease (COPD), and a respiratory tumor.
  • pathogens such as bacterial, viral, fungal, parasitic, atypical, community-acquired, healthcare-associated, hospital
  • the respiratory pathogens preferably include Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Stenotrophomonas maltophilia, Haemophilus parainfluenzae, Escherichia coli, Enterococcus faecalis, Serratia marcescens, Haemophilus parahaemolyticus, Enterococcus cloacae, Candida albicans, Moraxella catarrhalis, Streptococcus pneumoniae, Citrobacter freundii, Enterococcus faecium, Klebsiella oxytoca, Pseudomonas fluorescens, Neisseria meningitidis, Streptococcus pyogenes, Pneumocystis jirovecii, Klebsiella pneumonia
  • the hospitalized pneumonia is selected from the group consisting of hospitalized community-acquired pneumonia (hCAP), hospital-acquired (or nosocomial) pneumonia (HAP), ventilator-associated pneumonia (VAP), healthcare-associated pneumonia (HCAP), and severe community-acquired pneumonia (SCAP),
  • hCAP hospitalized community-acquired pneumonia
  • HAP hospital-acquired (or nosocomial) pneumonia
  • VAP ventilator-associated pneumonia
  • HCAP healthcare-associated pneumonia
  • SCAP severe community-acquired pneumonia
  • the implant or tissue infection is selected from the group consisting of burn wound infections, cardiology-associated infections, catheter-associated infections, deep skin and tissue infections diabetic foot infections, orthopedic implant infections, implant infections and surgical site infections,
  • the intra-abdominal infection is selected from the group consisting of acute abdomen, ascites, cholecystitis, diverticulitis, peritonitis, and surgical site infections,
  • the urinary tract infection is selected from the group consisting of catheter-associated urinary tract infection, complicated cystitis, urosepsis, and pyelonephritis.
  • inflammatory disease refers to a disease in which the immune system attacks and/or damages the body’s own tissues, resulting in an inflammation.
  • the inflammatory disease is selected from the group consisting of atherosclerosis, an autoimmune disease, allergy, asthma, a coeliac disease, glomerulonephritis, hepatitis, and an inflammatory bowel disease.
  • inflammation may be part of the normal healing process.
  • the substance (under test) may be related to an inflammation resulting from normal healing process or from an inflammatory disease.
  • sepsis refers to a life-threatening complication of a wide variety of infectious diseases. Sepsis arises when the body’s response to an infection causes injury to its own tissues and organs. Sepsis is usually caused by an inflammatory immune response triggered by the infection. Most commonly, the infection is bacterial, but it may also be fungal, viral, or protozoan. Disease severity partly determines the outcome. The risk of death from sepsis is as high as 30%, from severe sepsis as high as 50%, and from septic shock as high as 80%.
  • cancer disease refers to or describe the physiological condition in an individual that is typically characterized by unregulated cell growth.
  • cancer include, but are not limited to, lung cancer, preferably non-smallcell lung carcinoma (NSCLC) or small-cell lung carcinoma (SCLS), breast cancer, cervical cancer, gastric cancer, bladder cancer, skin cancer, nasopharyngeal cancer, neuroendocrine cancer, colon cancer, urothelial cancer, liver cancer, ovarian cancer, esophageal cancer, pancreatic cancer, kidney cancer, stomach cancer, esophageal cancer, renal cancer, head and neck cancer, brain cancer, lymphatic cancer, blood cancer, squamous cell cancer, laryngeal cancer, retina cancer, prostate cancer, uterine cancer, testicular cancer, bone cancer, lymphoma, and leukemia.
  • cancer also encompasses cancer metastases.
  • a substance specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), that is indicative of a microorganism.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • exemplary microorganisms include but are not limited to a bacterium, virus, fungus and protozoa.
  • the substance can be a pathogen.
  • Substances such as pathogens that can be handled, tested, or analysed with the cartridge of the present invention include, but are not limited to, Staphylococcus epidermidis, Escherichia coli, methicillin-resistant Staphylococcus aureus (MSRA), Staphylococcus aureus, Staphylococcus hominis, Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus capitis, Staphylococcus warneri, Klebsiella pneumoniae, Haemophilus influnzae, Staphylococcus simulans, Streptococcus pneumoniae and Candida albicans.
  • Staphylococcus epidermidis Escherichia coli, methicillin-resistant Staphylococcus aureus (MSRA), Staphylococcus aureus, Staphylococcus hominis, Enterococcus faecalis, Pse
  • Substances that can be handled, tested, or analyzed with the cartridge of the present invention also encompass substances responsible for a variety of sexually transmitted diseases selected from the following: gonorrhea (Neisseria gonorrhoeae), syphilis (Treponema pallidum), Chlamydia (Chlamydia trachomatis), nongonococcal urethritis (Ureaplasm urealyticum), yeast infection (Candida albicans), chancroid (Haemophilus ducreyi), trichomoniasis (Trichomonas vaginalis), genital herpes (HSV type I & II), HIV I, HIV II and hepatitis A, B, C, G, as well as hepatitis caused by TTV.
  • the substance comprises or consists of a nucleic acid molecule from the above microorganism.
  • the substance specifically the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), is a biomarker.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • biomarker refers to a biological molecule found in blood, other body fluids, or tissues that is an indicator of a normal or abnormal process, or of a condition or disease.
  • a biomarker may be used to foresee how well the body responds to a treatment for a disease or condition, or may be used to associate a certain disease or condition - or outcome of disease - to a certain value of said biomarker found in a sample e.g. a blood sample.
  • Biomarkers are also called molecular markers and signature molecules. If the biomarker is used to predict the probable course and outcome of a disease or condition, it may be called a prognostic biomarker. If the biomarker is used to diagnose a disease or condition, it may be called a diagnostic biomarker.
  • the substance specifically the substance comprising or consisting of a nucleic acid molecule, which is handled, tested, or analyzed with the cartridge of the invention can specifically be designated as analyte.
  • the at least one substance may be part of a fluid, e.g. a liquid or a gas.
  • the fluid may be a fluidic sample.
  • the fluidic sample may be a processed, non-processed, native (as removed from the body or from another source) or not yet processed fluidic sample.
  • the fluidic sample may be any medium which is suitable to accommodate the at least one substance.
  • the sample handled, tested, or analyzed with the cartridge of the invention can be of any origin or nature, for example of biological, chemical natural, synthetic, or semi-synthetic origin. The invention is, thus, not limited to any specific sample origin. Any sample suspected to contain the at least one substance, specifically the substance comprising or consisting of a nucleic acid molecule, can be used in conjunction with the cartridge of the invention.
  • the sample is a biological sample (a processed, non-processed, native or not yet processed biological sample). More preferably, the biological sample is a bodily sample (a processed, non-processed, native or not yet processed bodily fluid). Even more preferably, the bodily sample is a bodily fluid (a processed, non-processed, native or not yet processed bodily fluid) or bodily tissue (a processed, non-processed, native or not yet processed bodily tissue) sample. Specifically, the bodily tissue sample is a liquified bodily tissue sample.
  • a processed biological sample is based on/derived from a biological material.
  • the processed biological sample is a lysed sample or an extracted sample.
  • the biological sample as describe herein preferably may comprise cells and said cells may contain, in turn, a substance comprising or consisting of a nucleic acid molecule.
  • the biological sample may represent a culture medium or a culture supernatant, e.g. microbial culture medium, microbial growth medium.
  • the culture medium or the culture supernatant may comprise prokaryotes (bacteria, viruses) or eukaryotes, especially prokaryotic (bacteria, viruses) or eukaryotic pathogens.
  • the bodily sample such as bodily fluid or bodily tissue sample is incorporated directly into the cartridge of the present invention without further processing. Where desired or necessary, however, the bodily sample such as bodily fluid or bodily tissue sample, can be pre-treated before incorporation into the cartridge of the present invention.
  • the choice of pre-treatments will depend on the type of bodily sample such as bodily fluid or bodily tissue used sample and/or the nature of the substance.
  • the bodily fluid or bodily tissue can be concentrated via any conventional means to enrich the sub stance/ analyte.
  • Methods of concentrating the sub stance/ analyte include but are not limited to drying, evaporation, centrifugation, filtering, sedimentation, precipitation, and amplification.
  • the sub stance/ analyte is a nucleic acid molecule (e.g. DNA or RNA molecule), it can be extracted using various lytic enzymes or chemical solutions according to the procedures set forth in Sambrook et al.
  • sub stance/ analyte is a molecule present on or within a e.g. a cell having a cell nucleus or within an entity having no nucleus
  • extraction can be performed using lysing agents including but not limited to denaturing detergent such as SDS or non-denaturing detergent such as thesit, sodium deoxylate, triton X-100, and tween-20.
  • the bodily tissue sample may be, for example, liquefied before incorporation into the cartridge of the present invention.
  • bodily sample refers to any sample that is derived from the body of an individual. Especially, the term “bodily sample” refers to any sample that is derived from the body of an individual and comprises a substance, specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • a nucleic acid molecule e.g. DNA or RNA molecule.
  • the term “bodily sample” encompasses a bodily fluid sample and a bodily tissue sample.
  • tissue sample refers to any tissue sample that is derived from the body of an individual.
  • tissue sample refers to any sample that is derived from tissue of an individual and comprises a substance, specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • Said bodily tissue sample encompasses a skin flake, skin biopsy, hair follicle, biopsy tissue, tissue explant, and tissue section, the bodily tissue sample also encompasses tumor tissue sample.
  • Said bodily tissue sample may be removed from a patient or (control) subject by conventional biopsy techniques. It is preferred that the bodily tissue sample is a liquified bodily tissue sample.
  • bodily fluid sample refers to any fluidic sample that is derived from the body of an individual.
  • the term “bodily fluid sample” refers to any fluidic sample that is derived from fluid of an individual and comprises a substance, specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • a nucleic acid molecule e.g. DNA or RNA molecule.
  • any bodily fluid suspected to contain the at least one substance can be used in conjunction with the subject cartridge.
  • the body fluid sample may be a respiratory sample, a blood sample, an urine sample, a sputum sample, a breast milk sample, a cerebrospinal fluid (CSF) sample, cerumen (earwax) sample, a gastric juice sample, endolymph fluid sample, perilymph fluid sample, peritoneal fluid sample, pleural fluid sample, saliva sample, sebum (skin oil) sample, semen sample, sweat sample, tears sample, cheek swab, vaginal secretion sample, liquid biopsy, or vomit sample including components or fractions thereof.
  • CSF cerebrospinal fluid
  • cerumen earwax
  • the disease may be a respiratory disease.
  • the bodily sample such as a bodily fluid or bodily tissue sample is preferably taken for the purpose of a scientific test, such as for diagnosing a disease, e.g. a respiratory disease, for example, by detecting and/or identifying a pathogen or the presence of a tumor marker in a bodily sample which is preferably relevant for the diagnosis of a respiratory disease.
  • a bodily sample in the context of the present invention comprises cells, for example, pathogens or cells of the individual the bodily sample originated from, for example, tumor cells.
  • the preferred bodily samples are samples that are relevant for the diagnosis of a respiratory disease.
  • Such bodily samples may be respiratory samples, i.e. bodily samples derived from the respiratory tract, and non-respiratory samples, i.e. bodily samples that are not derived from the respiratory tract.
  • the respiratory tract in the context of the present invention preferably comprises the nose, nasal passages, paranasal sinuses, throat, pharynx, voice box, larynx, trachea, bronchi, bronchioles, and lungs, including respiratory bronchioles, alveolar ducts, alveolar sacs, and alveoli.
  • respiratory samples in the context of the present invention are sputum, pus (e.g., pus from the paranasal cavity), bronchial secretion, tracheal secretion, endotracheal secretion, bronchial aspirates, tracheal aspirates, endotracheal aspirates, bronchial lavage, bronchoalveolar lavage (BAL), bronchial swab, nasopharyngeal swab, laryngeal swab, and lung biopsies.
  • Preferred non-respiratory samples used in the present invention are relevant for the diagnosis of respiratory diseases.
  • non-respiratory samples in the context of the present invention are blood, pus, pleural fluid, pleural punctates, gastric juice, gastric aspirates, and drainages or punctate fluids from other body locations.
  • the term “individual”, as used herein, refers to any subject which may be analyzed or tested with the cartridge of the present invention.
  • the individual is preferably an animal, more preferably a mammalian animal including a human being.
  • an individual in the context of the present invention may be a mouse, rat, guinea-pig, rabbit, cat, dog, goat, sheep, pig, cow, horse, or human, preferably a human.
  • the individual may be a patient, wherein the term “patient” refers to an individual suffering from a disease or condition, or being suspected of suffering from a disease or condition.
  • molecular biological test refers to any molecular biological assay which allows the detection and/or analysis of at least one substance, specifically substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • the molecular biological test includes microbiological tests.
  • a preferred molecular biology test is a polymerase chain reaction (PCR) or a reverse transcription (RT) polymerase chain reaction (PCR).
  • biochemical test refers to any biochemical assay which allows the detection and/or analysis of at least one substance, specifically substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • nucleic acid molecule e.g. DNA or RNA molecule
  • the nucleic acid molecule may be comprised in a liquid such as a fluid or gas. It may be handled, tested, or analyzed with the cartridge of the present invention.
  • the nucleic acid molecule e.g. DNA or RNA molecule
  • the nucleic acid molecule may have to been extracted from its origin e.g. microorganism, and may have to be present in detectable amounts to be analysed. This may be achieved using nucleic acid molecule purification, extraction and amplification methods as described below.
  • the provided nucleic acid molecule may be attached to a probe which can be detected based on its color/fluorescence.
  • nucleotide refers to an organic molecule consisting of a nucleoside and a phosphate.
  • a nucleotide is composed of three subunit molecules: a nucleobase, a five-carbon sugar (ribose or deoxyribose), and a phosphate group consisting of one to three phosphates.
  • the four nucleobases in DNA are guanine, adenine, cytosine and thymine; in RNA, uracil is used in place of thymine.
  • the nucleotide serves as monomeric unit of nucleic acid molecules, such as deoxyribonucleotide acid (DNA) or ribonucleotide acid (RNA).
  • DNA deoxyribonucleotide acid
  • RNA ribonucleotide acid
  • the DNA molecule may be a double stranded DNA, a genomic DNA, or a complementary DNA (cDNA) molecule.
  • the RNA molecule may be a messenger RNA (mRNA), a small nucleolar RNA (snoRNAs), a ribosomal RNA (rRNA), or a transfer RNA (tRNA) molecule.
  • mRNA messenger RNA
  • snoRNAs small nucleolar RNA
  • rRNA ribosomal RNA
  • tRNA transfer RNA
  • nucleotide sequence or “polynucleotide” are interchangeably used herein and refer to single-stranded and double-stranded polymers of nucleotide monomers, including without limitation, 2'-deoxyribonucleotides (DNA) and ribonucleotides (RNA) linked by internucleotide phosphodiester bond linkages, or internucleotide analogs, and associated counter ions, e.g., H+, NH4+, trialkylammonium, Mg2+, Na+, and the like.
  • a nucleotide sequence or polynucleotide may be composed entirely of deoxyribonucleotides, entirely of ribonucleotides, or chimeric mixtures thereof and may include nucleotide analogs.
  • a “nucleic acid amplification method”, in the context of the present invention, is any molecular biological technique that is suitable for amplifying, i.e. multiplying, a nucleic acid, wherein the amplification may be linear or exponential.
  • nucleic acid amplification methods are polymerase chain reaction (PCR), nucleic acid sequence-based amplification (NASBA), ligase chain reaction (LCR), strand displacement amplification (SDA), multiple displacement amplification (MDA), Q-beta replicase amplification, and loop-mediated isothermal amplification.
  • the amplification method may be specific for a certain nucleic acid such as a specific gene or a fragment thereof, or may be universal such that all or a specific type of a nucleic acid, such as mRNA, is amplified universally.
  • the skilled person may design oligonucleotide primers which specifically hybridize to the nucleic acid of interest and use these primers in a PCR experiment.
  • a “nucleic acid analysis method” in the context of the present invention is any method that allows for detection and/or identification of a specific nucleic acid, wherein the term “detection” also comprises the quantitative determination of a nucleic acid.
  • the detection and/or identification may be based on specific amplification, for example, by the amplification of a specific DNA fragment using oligonucleotide primers specific for said DNA fragment in the polymerase chain reaction (PCR).
  • PCR polymerase chain reaction
  • the detection and/or identification may also be achieved without amplification, for example, by sequencing the nucleic acid to be analyzed or by sequence specific hybridization, for example, in the context of a microarray experiment. Sequencing techniques and microarraybased analysis are well known procedures in the field. The sequencing includes next generation sequencing.
  • the nucleic acid to be isolated, amplified, detected and/or identified may be DNA such as double stranded DNA, genomic DNA, or complementary DNA (cDNA).
  • the RNA may be messenger RNA (mRNA), small nucleolar RNA (snoRNAs), ribosomal RNA (rRNA), or transfer RNA (tRNA).
  • the nucleic acid amplification and/or analysis method is a polymerase chain reaction (PCR) or a reverse transcription polymerase chain reaction (RT-PCR).
  • PCR polymerase chain reaction
  • RT-PCR reverse transcription polymerase chain reaction
  • the PCR is selected from the group consisting of digital PCR, realtime PCR (quantitative PCR or qPCR), preferably TaqMan qPCR, multiplex PCR, nested PCR, high fidelity PR, fast PCR, hot start PCR, and GC-rich PCR.
  • the digital PCR may be digital droplet PCR or digital partition PCR.
  • PCR polymerase chain reaction
  • a PCR reaction may be carried out in a single test tube or chamber simply by mixing DNA (deoxyribonucleic acid) with a set of reagents and performing thermal cycles. Thereby, the following steps are repeated several times thereby doubling the number of double DNA strands in each cycle:
  • DNA polymerase thermoostable polymerase
  • PCR reverse-transcriptase-PCR
  • qRT-PCR or RT-qPCR real time quantitative PCR
  • inverse PCR irt-PCR
  • immune PCR immune PCR
  • agglutination-PCR e.g. PCR-Test (reverse-transcriptase-PCR), qRT-PCR or RT-qPCR (real time quantitative PCR), inverse PCR, irt-PCR (immunoquantitative real time PCR), immune PCR, agglutination-PCR.
  • RT-PCR reverse transcription polymerase chain reaction
  • a master mix For a PCR reaction, a master mix is used.
  • the master mix contains dNTPs (deoxyribose nucleoside triphosphate), e.g.
  • dATP deoxyadenosine triphosphate
  • dGTP deoxyguanosine TP
  • dTTP deoxythymidine TP
  • dCTP deoxy cytidine TP
  • Taq DNA polymerase enzymes MgCh, stabilizers, and enhancers in a reaction buffer.
  • Master mix, specific primers and/or universal primers and probes for the detection/amplification of the at least one substance and the at least one substance have to be brought together to perform the PCR reaction.
  • the term “probe”, as used herein, may refer to detectable “auxiliary” molecules or other materials, e.g. using optical detection or optical detection units.
  • the probe may comprise a Taqman probe.
  • a Taqman probe is a hydrolysis probe that is designed to increase the specificity of quantitative PCR.
  • the probe may have fluorescence characteristics in order to ease or to admit optical detection.
  • double marked probes with quenchers may be used, especially in order to enhance the substances under tests that may be tested within one chamber.
  • only one color may be detected using e.g. intercalating dyes or other dyes.
  • the PCR system may have applications in a broad range of molecular biology and biotech lab experiments, including cloning (or synthesis of specific DNA fragments), sequencing, genotyping, nucleic acid synthesis, gene expression, generation of NGS (next generation sequencing) libraries, and mutagenesis.
  • a PCR master mix may specifically help researchers and scientists to enhance their PCR assay performance by providing a spectrum of benefits, including saving time and reducing the chances of any errors/cross-contamination in preparing PCR formulations. They are often utilized in routine or high-yielding PCR.
  • PCR master mixes are available in liquid and lyophilized forms.
  • the liquid form mix is required to be stored at a temperature between -20°C to +4°C and is typically cheaper than the lyophilized or freeze-dried mixes.
  • Lyophilized PCR master mixes can be stored at ambient temperatures for a longer period. Moreover, they are easy to transport and while running PCR, the solution only needs to be reconstituted in the buffer solution, which comes with the master mix.
  • Master mixes for real-time PCR may further include at least one probe, especially a fluorescent compound and/or fluorescence enhancing compound and/or fluorescence suppression compound or molecule.
  • a probe especially a fluorescent compound and/or fluorescence enhancing compound and/or fluorescence suppression compound or molecule.
  • only one dye may be used within one chamber, e.g. an intercalating dye.
  • the probe may be provided separate from the master mix, e.g. within a detection chamber, e.g. within the (PCR) disc wheel mentioned below.
  • processing in the context of the present invention refers in general to every treatment that comprises a change in one or more physical properties of the fluid such as fluidic sample being processed when said physical property/properties is/are determined before and after the treatment/processing.
  • the “processing” in the sense of the present invention may comprise the liquefaction of the liquid such as fluidic sample so that the viscosity of the liquid such as fluidic sample is reduced by the processing procedure.
  • the “processing” may comprise (in addition or alternatively) the lysis of the sample, meaning the liquefaction and disintegration of pathogens present in the sample.
  • Such cells may be prokaryotic (e.g. no nucleus) or eukaryotic cells (e.g.
  • nucleus comprising DNA for example, bacterial cells, yeast cells, fungal cells, animal cells, mammalian cells etc., wherein processing may lead to liquefaction and/or lysis of all cells or only of a specific type of cells or a subset of bacteria.
  • an “automated process”, in the context of the present invention, means a process which is operated and/or controlled by automation.
  • an “automated process”, in the context of the present invention does not require or include any manual handling steps.
  • method steps that are performed in an automated process are preferably performed by an apparatus or device, such as the cartridge as described herein, which is preferably programmable to perform said method steps in a sequential order.
  • lysis buffer refers to a buffer that is suitable for lysis of cells with the purpose of analyzing the contents of the cells, such as analyzing the nucleic acid molecules contained in the cells.
  • the lysis buffer according to the present invention is suitable for the lysis of mammalian cells and/or microbial cells, such as bacterial and yeast cells.
  • the lysis buffer of the present invention is preferably suitable for the lysis of bacteria of a family selected from the group consisting of Mycobacteriaceae, Pseudomonadaceae, Mycoplasmataceae, Chlamydiaceae, Enter obacteriaceae, Staphylococcaceae, Streptococcaceae,
  • Xantomonadaceae Moraxellaceae, Legionellaceae, Burkholderiaceae.
  • Corynebacteriaceae, Neisseriaceae, Bacteroides, and Pasteurellaceae and/or for the lysis of yeasts of a family selected from the group consisting of Saccharomycetaceae, Sporidiobolaceae, Trichocomaceae, and Pneumocystidaceae .
  • the lysing of cells may additionally or alternatively be achieved by applying osmotic and/or mechanical disruption of cells.
  • cell lysis refers to a technique that destroys and/or disrupts cells for the purpose of analyzing the contents of the cells, such as analyzing the at least one substance (e.g. nucleic acid molecule such as DNA or RNA molecule) contained in the cells.
  • the cells may be mammalian cells and/or microbial cells, such as bacterial and yeast cells.
  • lysate refers to the product of enzymatic, osmotic, and/or mechanical disruption of the cell membranes of a cell population.
  • Cell lysates are widely used for the isolation of cellular components such as ucleic acid molecules like DNA, RNA, proteins or whole organelles.
  • a lysate is produced in order to detect, analyze, and/or quantify the at least one substance, specifically the substance comprising or consisting of a nucleic acid molecule comprised in the cell population.
  • the term “elution buffer”, as used herein, refers to a buffer solution which allows the removal of compounds from a substrate to which the compounds are bound (e.g. via covalent and/or non-covalent bounds).
  • the elution buffer allows the removal of the at least one substance, specifically the at least one substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), from nucleic acid binding beads after they have been captured on said beads.
  • the elution buffer is designed to optimally remove the nucleic acid material from the beads by simply adding the elution buffer to the beads. The removed nucleic acid molecules are finally dissolved in the elution buffer and can be further processed.
  • eluate refers to a discharged mixture of solvents and dissolved substances (also designated as “wash out”).
  • the mixture of solvents and dissolved substances eluted or recovered from beads e.g. magnetic beads
  • the mixture of elution buffer and dissolved substance specifically the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule) can be designated as eluate.
  • biological filter indicates any type of filter that prevents that living cells are released into the environment, e.g. bacteria, viruses, etc.
  • a cartridge for handling at least one substance specifically at least one substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), is provided, comprising:
  • a stationary unit comprising a stationary fluidic system
  • a movable unit comprising a movable fluidic system, wherein the movable unit is movable relative to the stationary unit.
  • the relative movement may be a movement along at least one movement direction or along at least two movement directions, e.g. a back and forth movement or a clockwise versus counter clockwise rotation.
  • the relative movement may be a rotation e.g. in order to provide e.g. a switching wheel that is able to connect one pump port with a plurality of channels of the stationary fluidic system depending on a switching position within a comparably large switching angle, e.g. almost one quadrant, one quadrant or more than one quadrant of a circle or at least an angle within the range of 30 degrees to 80 degrees.
  • the angle may be less than 180 degrees or less than 120 degrees.
  • the angle between a first switching position and a second switching portion may be different from a 180 degree angle.
  • only one pump port e.g. comprised within the stationary fluidic system may be connected to other parts of the stationary fluidic system using the movable fluidic system.
  • two pump ports may be connected to e.g. a plurality of channels of the stationary fluidic system depending on a switching position within the comparably large switching angle, e.g. to pairs of channels that are connected with a reaction chamber and or a buffer chamber/compartment within the stationary fluidic system.
  • the movable system may comprise at least two channels for both flow direction, e.g. from the pump and back to the pump.
  • the stationary fluidic system and the movable fluidic system may be provided to receive, to guide, and/or to retain a fluid.
  • the fluid may comprise or consist of at least one substance, e.g. a biological substance, such as a microbiological substance.
  • the fluid may comprise chemicals, preferably fluids to treat and/or to prepare the at least one substance for a test.
  • the at least one substance may be a substance as mentioned above in the introductory part of the description. Specifically, the at least one substance may comprise or may consists of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • a substance specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), which is associated with a particular disease or condition or with a specific disease or condition stage.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the at least one substance may be associated with infectious diseases, inflammatory diseases, sepsis, autoimmune diseases, cancer diseases (or simply cancer), or any combinations thereof.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the infectious disease may be a respiratory disease such as pneumonia like hospitalized pneumonia, an implant or tissue infection, an intra-abdominal infection, or a urinary tract infection.
  • a respiratory disease such as pneumonia like hospitalized pneumonia, an implant or tissue infection, an intra-abdominal infection, or a urinary tract infection.
  • the at least one substance may be indicative of a microorganism.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the at least one substance can be a pathogen.
  • the substance specifically the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), may be a biomarker.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the fluid may be a liquid or a gas.
  • the fluid may be a fluidic sample.
  • the fluidic sample may be a processed, non-processed or not yet processed fluidic sample.
  • the sample is a biological sample (a processed, non-processed or not yet processed biological sample).
  • the biological sample is a bodily sample (a processed, non-processed or not yet processed bodily fluid).
  • the bodily sample may be a bodily fluid (a processed, non-processed or not yet processed bodily fluid) or bodily tissue (a processed, non-processed or not yet processed bodily tissue) sample.
  • the bodily tissue sample may be a liquified bodily tissue sample.
  • the biological sample may also be a culture medium, e.g. a cell culture medium or a culture supernatant, e.g. a cell culture supernatant.
  • the at least one substance e.g. nucleic acid molecule such as DNA or RNA molecule
  • the at least one substance which is detected with the radiation transfer unit (RU) of the present invention is preferably labelled with a detectable dye specifically fluorescence marker/probe such as fluorophore.
  • the nucleic acid molecule such as DNA or RNA molecule as the at least one substance is labelled with a detectable dye specifically fluorescence marker/probe such as TaqMan probe.
  • a TaqMan probe is a hydrolysis probe that is designed to increase the specificity of quantitative PCR.
  • the at least one substance is part of a sample such as biological sample as described herein. If the sample contains cellular material and the at least one substance (e.g. nucleic acid molecule such as DNA or RNA molecule) is contained therein, the cellular material needs to be lysed first in order to release the at least one substance (e.g. nucleic acid molecule such as DNA or RNA molecule) from the cells. Subsequently, the at least one substance (e.g. nucleic acid molecule such as DNA or RNA molecule) is isolated from the cell debris and then purified. In case the sample already contains the at least one substance (e.g.
  • nucleic acid molecule such as DNA or RNA molecule
  • a PCR reaction is preformed to amplify the nucleic acid molecule such as DNA or RNA molecule before detection.
  • the PCR reaction is preferably conducted in the presence of a TaqMan probe.
  • the RNA is first transcribed into cDNA before the amplification reaction is performed.
  • a TaqMan probe consists of a fluorophore covalently attached to the 5’-end of the oligonucleotide probe and a quencher at the 3 ’-end.
  • fluorophores e.g. 6- carboxyfluorescein, acronym: FAM, or tetrachlorofluorescein, acronym: TET
  • quenchers e.g. tetramethylrhodamine, acronym: TAMRA
  • the quencher molecule quenches the fluorescence emitted by the fluorophore when excited by the cycler’s light source via Forster resonance energy transfer (FRET).
  • TaqMan probes are designed such that they anneal within a nucleic acid such as DNA region amplified by a specific set of primers.
  • TaqMan probes can be conjugated to a minor groove binder (MGB) moiety, dihydrocyclopyrroloindole tripeptide (DPI3), in order to increase its binding affinity to the target sequence; MGB-conjugated probes have a higher melting temperature (T m ) due to increased stabilization of van der Waals forces.
  • MGB minor groove binder
  • DPI3 dihydrocyclopyrroloindole tripeptide
  • the 5' to 3' exonuclease activity of the Taq polymerase degrades the probe that has annealed to the template. Degradation of the probe releases the fluorophore from it and breaks the proximity to the quencher, thus, relieving the quenching effect and allowing fluorescence of the fluorophore.
  • fluorescence detected in the quantitative PCR thermal cycler is directly proportional to the fluorophore released and the amount of nucleic acid such as DNA template present in the PCR. This signal can then be detected with the radiation transfer unit (RU) of the present invention.
  • At least one of the stationary fluidic system and the movable fluidic system may comprise a coupling portion of the stationary fluidic system or of the movable fluidic system.
  • the coupling portion may be comprised within the stationary fluidic system.
  • the coupling portion may comprise a channel that extends along the movable unit, e.g. along a distance or circumferential distance that equals the large switching angle or that corresponds to it.
  • the coupling portion may comprise a central chamber and at least two channels or a plurality of channels that are connected to the central chamber.
  • the free ends of the channels of the coupling portion may be arranges within a region that extends along the movable unit, e.g. along a distance or circumferential distance that equals the large switching angle or that corresponds to it.
  • the coupling portion may comprise a fluidic passage, an intermediate volume and at least two fluidic interface portions.
  • the fluidic passage may be an inflow opening or an outflow opening of the coupling portion depending on a flow direction.
  • the fluidic passage may be directly coupled to a pump port.
  • the intermediate volume may be a volume of an elongated channel or of a central portion of the coupling portion.
  • the cartridge in at least two different relative positions of the movable unit relative to the stationary unit, may be configured to establish a fluidic communication between the fluidic passage, e.g. comprised within the stationary fluidic system, and a lumen, preferably a channel, of the fluidic system to which the coupling portion does not belong, e.g. the movable fluidic system, via a fluidic flow passing first through the fluidic passage, then through the intermediate volume and thereafter through a respective one of the at least two fluidic interface portions or vice versa.
  • the fluidic passage e.g. comprised within the stationary fluidic system
  • a lumen preferably a channel
  • At least three different relative positions may be provided in which the cartridge is configured to establish the fluidic communication between the passage of the coupling portion and the fluidic system to which the coupling portion does not belong, e.g. is not part of.
  • the number of relative positions may be in the range of 2 to 15 or in the range of 3 to 10 allowing e.g. to select the same number of buffers and/or of reaction chambers of the first fluidic system.
  • a second coupling portion may be provided to enhance the switching angle to almost two quadrants to two quadrants or to more than two quadrants, e.g. allowing the selection of twice the number of compartments as mentioned above, e.g. in the range of four to 30 or in the range of 6 to 20.
  • about two quadrants of the circumference of a rotatable fluidic system may be used for the coupling portions and about two other quadrants may be used to arrange the channels of e.g. the stationary fluidic system connected to reaction chambers, buffers, etc. of e.g. the stationary fluidic system.
  • This may allow switching between at least 10 or at least 12 entities (reaction chamber, buffer, etc.) of the fluidic system of the cartridge allowing to perform sophisticated tests, e.g. nucleic acid amplification, nucleic acid quantification, and/or nucleic acid analysis tests such as PCR or RT-PCR tests within a micro reactor.
  • the fluidic system of the micro reactor may be formed by the fluidic system of the cartridge.
  • the fluidic system of the micro reactor is a separate part that is separate from other components of the micro reactor.
  • the cartridge may be disposable. No cleaning of the other parts of the micro reactor may be necessary after performing a test.
  • the maximum lateral dimension of the cartridge may be less than 20 cm (centimeter) and e.g. more than 7 cm.
  • the other lateral dimension of the cartridge may be less than 15 cm and e.g. more than 5 cm.
  • the thickness of the cartridge may be below 2 cm and e.g. more than 5 mm (millimeter).
  • the whole fluidic system of the micro reactor may be implemented using a small cartridge.
  • the at least two fluidic interface portions or the at least three interface portions may have a distance matched to at least two relative positions of the movable unit relative to the stationary unit.
  • the at least two interface portions may be implemented as taps that tap from the intermediate volume of the coupling portion and/or that may be comprised within the coupling portion.
  • the taps of one coupling portion may form a group of taps.
  • a further group of taps may correspond to the selectable switching positions on the respective coupling portion but may be arranged on the side of other compartments, e.g. reaction chamber(s), buffers, etc., preferably of the stationary fluidic system.
  • the number of taps on the side of the entities/compartments of the e.g. stationary fluidic system, e.g. reaction chamber(s), buffers, etc. may be larger, e.g. about or exactly twice as large as the number of taps on the side of the respective coupling portion. This may be possible by using different offsets between the taps of the coupling portion and the taps on the other side of the fluidic system to which the coupling portion belongs. Thus, the offset of the taps of the coupling portion may be twice as large as the taps on the other side other side of the fluidic system to which the coupling portion belongs.
  • the taps on the other side may form a further group of taps.
  • the smaller offset may be in the range of 1 mm to 4 mm, e.g. a value about 2.5 mm or of exactly 2.5 mm may be used.
  • the larger offset may be in the range of 3 mm to 7 mm or in the range of 4 mm to 6 mm, e.g. a value about 2.5 mm or of exactly 2.5 mm may be used.
  • the coupling portion in the at least two different relative positions or in the at least three different positions, may be configured to provide fluidic communication between the fluidic passage and the fluidic system to which the coupling portion does not belong using a respective one of the at least two fluidic interface portions and at least one counter fluidic interface portion comprised in the fluidic system to which the coupling portion does not belong.
  • the fluidic interface related to the coupling portion may comprise several fluidic sub-interfaces.
  • a fluidic interface may be formed by a preferably plane surface area at the border between a stationary interface portion and a movable, especially rotatable interface portion.
  • At least one gasket may be part of the fluidic interface in order to provide fluid tight connection between the interface portion and the counter interface.
  • the gasket may have a further sealing interface on the side that is opposite to the side facing the fluidic interface portion.
  • the further sealing interface may be stationary, e.g. not involving movable parts or entities.
  • the sealing may be formed around the end of channels that are adjacent to the gasket and that are preferably aligned to a respective through hole of the gasket. The same may be valid for the fluidic sub-interfaces.
  • the fluidic (sub-)interface may be formed in a specific switching position, e.g. when an interface portion is aligned to the counter interface portion.
  • both portions may be aligned e.g. at the same x position and the same y position and only the vertical position may be different, e.g. the z position.
  • Axial portions of the interface portion and/or of the counter interface portion may extend in the z direction.
  • An axial gasket may be used to provide fluid tightness via a comparably large temperature range and/or between parts of different temperature, e.g. compared to a radial gasket.
  • the at least two fluidic interface portions or the at least three fluidic interface portions may be discrete/separate portions, e.g. there may be intermediate spaces between the interface portions.
  • the interface portions may be formed using taps.
  • the intermediate spaces may be the spaces between “adjacent” or neighboring taps.
  • the discrete fluidic interface portions may allow more fluid tightness and/or more exact positioning at the switching positions.
  • the at least two fluidic interface portions or the at least three fluidic interface portions may be at least two different sub-portions of a continuous fluidic interface portion. Although a continuous fluidic interface portion may be used, the switching positions may still be discrete, e.g. at only some predefined angles.
  • a continuous interface portion may be simpler, e.g. compared to the usage of discrete interface taps.
  • the passage may be connected to a first flow port connected to a pump or to a part of the pump, preferably to a flexible hose that is configured to be pressed and released by a peristaltic pump.
  • the peristaltic pump may comprise at least one roller or wheel.
  • at least one gear may be used. The tooth of the at least one gear may allow to compress the flexible hose more tightly, e.g. compared to a roller resulting in better pumping results, especially within a pumping mode for low volumes.
  • Low volumes may be less than 50 microliters, less than 40 microliters, less than 30 microliters but more than 1 microliter or more than 2 microliters to give only an example for a lower range.
  • the first flow port may be an inflow opening or an outflow opening for fluid into or out of fluidic system formed by the first fluidic system and by the second fluidic system, e.g. depending on the flow direction of the flow driven by the pump.
  • the movable unit may be used to connect e.g. a specific reaction chamber and/or a specific buffer chamber preferably of the first fluidic system with a pump, e.g. with the flexible hose of a peristaltic pump such that a closed fluidic loop is established.
  • the closed fluidic loop may allow to pump fluid out of the chamber/buffer or into the chamber/buffer.
  • the fluid that is pumped out of the chamber/buffer or that has to be pumped into the chamber/buffer may be stored preliminary within a channel of the movable unit, e.g. within an enlarged channel comprising e.g. at least one winding or at least two windings or within an auxiliary storage chamber of the movable/rotatable fluidic system.
  • the movable unit may not comprise a long-term storage buffer chamber or a reaction chamber itself.
  • the movable unit may have a simple construction and/or the machine used to perform the test.
  • Both units e.g. the stationary unit and the movable unit may be part of the same cartridge.
  • the cartridge may comprise only one movable unit, especially a movable unit for switching purposes.
  • the cartridge may comprise at least one further movable unit, e.g. a disc wheel comprising at least two chambers for performing temperature cycles, e.g. as part of a nucleic acid amplification, nucleic acid quantification, and/or nucleic acid analysis test such as PCR or RT-PCR test.
  • the cartridge may be configured to be inserted into a device (system) or machine that comprises auxiliary units, e.g. at least one heating unit and/or at least one stirring unit and/or at least one detection unit etc., see the description of the figures below for further details.
  • the substance may be a biological substance such as microbiological substance, e.g. as defined above.
  • the substance comprises or consists of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • the health risk for personal inherent to the processing of material possibly infected with viruses or bacteria may remain limited essentially to the first step in which the sample is placed in the sample tube.
  • the risk of contamination may be very low because the whole procedure can be carried out in a single cartridge comprising at least one reaction chamber. After the optional lysing step pathogens may be effectively inactivated.
  • the at least two relative positions may be relative positions of a first group of relative positions.
  • the relative positions of the first group of relative positions may be different from relative switching positions of a second group mentioned below.
  • the first fluidic system may comprise at least two fluidic compartments, e.g. reaction chambers and/or liquid buffers or other buffers. Each of the at least two fluidic compartments may be connected to at least two fluidic channels of the first fluidic system in order to allow internal filling and or internal removal of chemicals from the respective fluidic compartment.
  • the fluidic passage of the coupling portion may be fluidically connected to at least one pump port.
  • the pump port may be configured to be connected to a pump unit, e.g. to the flexible hose of a peristaltic pump or to another type of pump.
  • the fluidic passage may be fluidically connected with a first fluidic compartment of the at least two fluidic compartments via a first fluidic interface portion of the at least two fluidic interface portions and via at least a first fluidic channel of the at least two fluidic channels.
  • the first fluidic compartment may be connected via at least a second fluidic channel of the at least two fluidic channels to the pump unit in order to create a closed fluidic loop.
  • a closed loop from one pump port to another pump port may be used thereby it may be possible to use pressure as well as suction for fluid transport. This may provide redundancy under the following circumstances, e.g. if the overall fluidic system comprises a hole or other leakage.
  • a portion of liquid may be transported that is bordered by a first portion of a gas, e.g. air on one end and by a second portion of a gas, e.g. air on the other end.
  • a gas e.g. air on one end
  • a second portion of a gas e.g. air on the other end.
  • the first fluidic compartment may be connected via at least a second fluidic channel of the at least two fluidic channels to another outlet or inlet of the first fluidic system or of the second fluidic system in order to create a closed fluid loop.
  • both connections to and from the pump may have portions within the movable unit. This may allow the most sophisticated switching solutions.
  • the coupling portion may be a first coupling portion.
  • a second coupling portion may be used that is similar to the first coupling portion, e.g. as part of same fluid system as first coupling portion, see below.
  • the second coupling portion may be used to provide a fluidic connection to the second pump port or to the fluid outlet.
  • the other pump connection may be completely part of the first fluid system, i.e. no movable unit or movable fluid system may be involved within this fluidic connection.
  • the number of fluidic compartments may be in the range of two to 15 or in the range of 3 to 15.
  • At least one of the fluidic compartments or at least two of the fluidic compartments may be storage chamber(s) that may store a chemical or a chemical product, e.g. ethanol, water, isopropanol, washing solution, etc.
  • a chemical or a chemical product e.g. ethanol, water, isopropanol, washing solution, etc.
  • At least one of the fluidic compartment or at least two of the fluidic compartments may be reaction chamber(s).
  • At least one of the reaction chamber or at least two of the reaction chambers may comprise stirring elements, e.g. an agitator, e.g. an agitator that is rotatable and/or movable using an external magnet.
  • the fluidic passage in a second relative position of the at least two relative positions, may be fluidically connected with a second fluidic compartment of the at least two fluidic compartments via the second fluidic interface and via at least a third fluidic channel of the at least two fluidic channels.
  • the second fluidic compartment may be connected via at least a fourth fluidic channel of the at least two fluidic channels to the pump unit in order to create a second closed fluidic loop.
  • the second closed fluid loop may be created after or before establishing the first closed fluid loop.
  • the direction of fluid transport may be chosen as is appropriate in both closed fluid loops, e.g. by switching the direction of a pump, e.g. of the movable part of a peristaltic pump.
  • the movement may be a rotation movement.
  • the coupling portion may comprise an elongated channel that extends along a circumferential direction relative to a rotation axis of the movable unit.
  • the channel in case of a translational relative movement, may extend along a straight movement direction of the movable unit.
  • the elongated channel may comprise at least two lateral fluidic connections that are configured to be connected to the at least two fluidic interface portions.
  • a first circumferential distance or straight distance may be used between the at least two lateral fluidic connect! ons/taps.
  • the first (circumferential) distance or offset may be twice as much as the (circumferential) distance between an input port to a chamber and an output port of the same chamber on the first unit, e.g. on the stationary unit.
  • This choice of the distances or offsets may have several technical effects, e.g. dense arrangement of switching positions and/or providing enough spare between the taps of the coupling portion in order to close channel openings in the neighborhood of a main channel opening that is connected to a tap of the coupling unit.
  • the elongated channel may be a first elongated channel.
  • a second elongated channel is mentioned below. As already mentioned above, this may allow extension of the switching range, e.g. over a region of about two quadrants of a circle that comprises all switching positions and/or enabling connection of both pump ports via the movable unit and via the stationary unit.
  • the coupling portion may be used, e.g. a more central chamber comprising the fluidic passage (inlet or outlet) and several channels extending from the more central chamber to the fluidic interface portions.
  • the channels mentioned above may be arranged within the cartridge, i.e. within stationary part or movable part.
  • the channels may be protected and/or no additional parts may be necessary for switching, especially no flexible parts.
  • usage of injection molding or of other micromachining technology is possible as well, e.g. in order to enable cost effective mass production, e.g. more than 1000 cartridges or more than 10000 cartridges may be produced.
  • the movement may be a rotation movement.
  • a rotation angle between a first relative position of the at least two relative positions and a second relative position of the least one two relative positions may have a value in the range of 30 degrees to 110 degrees, in the range of 50 degrees to 100 degrees or in the range of 60 degrees to 90 degrees.
  • a considerable large range may be covered and used for several switching positions and/or e.g. for connecting several compartments to a pump port. This may be enabled by the usage of the fluidic coupling portion/fluidic coupling system.
  • the movement may be a rotation movement.
  • the cartridge may comprise an axial gasket between the stationary unit and the movable unit.
  • the axial gasket may form part of a first interface (sub-interface).
  • the axial gasket may comprise at least one hole that extends in the direction of the rotation axis of the movable unit.
  • the fluidic connection formed by the first interface (sub-interface) may comprise a fluidic and e.g. mechanical connection (mechanical pressure, physical contact) of the first unit (stationary), then a connection formed by a through hole of the gasket, and then a fluidic and mechanical connection (e.g. movable, rotatable between switching positions but stationary if the switching position is selected, e.g. mechanical pressure, physical contact) to the second unit.
  • the axial gasket may be arranged in a circular groove formed in the first unit, e.g. in the stationary unit.
  • the groove may have a structured outer wall, preferably an outer wall comprising axially extending protrusion and axially recessed recesses in alternate arrangement.
  • rotation of the gasket during rotation of the movable (rotatable) unit may be prevented in an easy way.
  • the groove may have a circular inner wall, preferably without axially extending protrusion and/or axially recessed recesses.
  • the axial gasket may provide a more fluid tight connection, e.g.
  • temperature may not or may rarely influence tightness of fluidic interfaces at the axial gasket, especially of the first interface or the first sub-interface.
  • Axial openings may be provided on the second unit that connect a first channel and other channels of the second unit (movable/rotatable unit) in axial direction to the respective one of the at least two interface portions of the stationary fluidic system.
  • An axial gasket or the axial gasket mentioned above may form a sealing around these axial holes in a respective switching position.
  • the channels of the fluidic systems may have end portions, e.g. taps or axial portions, that are curved or angled, e.g. by an angle in the range of 60 degrees to 120 degrees, 80 degrees to 100 degrees, or 90 degrees in order to allow usage of the axial gasket.
  • the movable unit may comprise a first channel.
  • the first channel may comprise a first end portion and a second end portion. In the at least two different relative positions, the first end portion may be arranged spatially and/or fluidically closer to the at least two interface portions.
  • the second end portion may be arranged away from the at least two interface portions, preferably on an opposite side of the movable unit compared to the side on which the first end portion is arranged.
  • the first channel may be the main channel of the movable unit, e.g. comprising the largest volume.
  • the first channel may be used as a preliminary storage for fluid, e.g. fluid removed from or to be filled in one of the buffers of the first fluidic system or for fluid to be filled in or to be removed from at least one reaction chamber of the first fluidic system, e.g. the stationary fluidic system.
  • the first end portion of the first channel or the second end portion of the first channel may be configured to be connected to other channels of the first fluidic system, e.g. via a further fluidic interface.
  • the coupling portion may be a first coupling portion
  • the fluidic passage may be a first fluidic passage and the intermediate volume may be a first intermediate volume.
  • the lumen/channel may be a first channel, preferably of the movable fluidic system.
  • the at least two fluidic interface portions may be interface portions of a first group.
  • At least one of the stationary fluidic system and the movable fluidic system may comprise a second coupling portion of the stationary fluidic system or of the movable fluidic system.
  • the second coupling portion may comprise a second fluidic passage, a second intermediate portion and at least two fluidic interface portions of a second group.
  • the cartridge may be configured to establish a fluidic communication between the second fluidic passage and a second channel of the fluidic system to which the second coupling portion does not belong via a fluidic flow passing first through the second fluidic passage, then through the second intermediate volume and thereafter through a respective one of the at least two fluidic interface portions of the second group or vice versa.
  • the second coupling portion may not only enhance the switching range, e.g. the switching angle considerably, especially within a second quadrant but may alternatively or additionally be used also as a backflow portion in the at least two different relative positions.
  • the at least two different relative positions may be positions of a first group that may be different from a second group of switching positions mentioned below.
  • the lumen/channel may be a first channel, preferably of the movable fluidic system,
  • the at least two different relative positions may be positions of a first group
  • the second coupling portion may be used for coupling to the first channel, especially independent whether there is a second channel, e.g. of the movable fluidic system, in a second group of relative positions that may be different from the first group, e.g. with no overlap of relative positions or of witching positions between the first group and the second group.
  • a second channel e.g. of the movable fluidic system
  • the switching range may be extended by the second coupling portion.
  • the second coupling portion may have at least one further function in addition to range extension which would also be possible by a respective extension of the first coupling portion.
  • the further function may e.g. relate to providing a backflow and/or to provide at least one other function, e.g. in combination with further channels of the fluidic system to which the coupling portion does not belong, e.g. of the movable fluidic system.
  • the at least two fluidic interface portions of the first group and/or the at least two fluidic interface portions of the second group may have a distance matched to the at least two relative positions of the movable unit relative to the stationary unit.
  • the second coupling portion may be configured to provide fluidic communication between the second fluidic passage and the fluidic system to which the coupling portion does not belong using the second group of fluidic interfaces each comprising a respective one of the fluidic interface portions and at least one counter fluidic interface portion comprised in the fluidic system to which the coupling portion does not belong.
  • the at least two relative positions may be relative positions of the first group of relative positions.
  • This second fluid interface may be that the rotation angle at which second flow port may be used to establish fluidic connections is enlarged.
  • the second passage may be connected directly to a pump or pump port or to another inlet or outlet of the first fluidic system.
  • the second flow port may be connected to a pump or to a part of the pump, preferably to a flexible hose that may be configured to be pressed and released by a driving unit of the peristaltic pump.
  • the second fluidic system may comprise a second channel.
  • the second channel of the second fluidic system may comprise a first end portion and a second end portion. In the at least two different relative positions, the first end portion of the second channel may be connected to the at least two interface portions of the second group.
  • the second end portion of the second channel of the second fluidic system may be arranged away from the at least two interface portions, preferably arranged circumferentially between the first end portion and an opposite side of the movable unit/rotatable unit compared to a side on which the first end portion is arranged.
  • the second coupling portion may be similar to the first coupling portion, preferably comprising a second elongated channel that may extend along a circumferential direction relative to a rotation axis of the movable unit or that may extend along a straight movement direction of the movable unit.
  • the second elongated channel may comprise at least two lateral fluidic connections that may be configured to be connected to the at least two fluidic interface portions of the second group.
  • a first circumferential offset between the at least two lateral fluidic connections may be twice as large as the offset between an input port to a chamber and an output port of the same chamber, e.g. in the stationary fluidic system.
  • the same values for the offsets may be used as mentioned above.
  • An angle between a first end of the second channel, a fulcrum of the rotatable second fluidic system and a second end of the second channel may be within the range of 30 degrees to 110 degrees, in the range of 50 degrees to 100 degrees or in the range of 60 degrees to 90 degrees. Thus, optimal switching capabilities may be provided.
  • the at least two relative positions may be relative positions of a first group of relative positions.
  • the lumen/channel may be a first channel, preferably of the movable fluidic system.
  • a fluidic connection may be established between the first fluidic passage and the first channel of the fluidic system to which the coupling portion does not belong, preferably of the movable fluidic system.
  • the cartridge may be configured to establish a fluidic communication between the fluidic passage and a second channel of the fluidic system to which the coupling portion does not belong via a fluidic flow passing first through the fluidic passage then through the intermediate volume and thereafter through a respective one of the at least two fluidic interface portions or vice versa.
  • the first channel preferably of the movable fluidic system, may be different from the second channel, preferably of the movable fluidic system.
  • the switching range may be extended by usage of the second channel of the fluidic system to which the coupling portion does not belong, preferably of the movable fluidic system.
  • claims 7 and claim 8 may allow sophisticated and/or advanced applications using small cartridges for performing tests involving several chemicals.
  • the first fluidic system may comprise at least one reaction chamber or at least two reaction chambers.
  • the at least one reaction chamber may be configured or the at least two reaction chambers may be configured to be heated by an external heating unit.
  • the at least one reaction chamber may comprise a stirring element or the at least two reaction chambers may comprise a respective stirring element that may be moved by an external stirring unit.
  • the fluidic system of the micro reactor may be appropriate to perform different kind of tests and/or to perform tests comprising a plurality of steps, e.g. more than 10 steps, more than 50 steps or more than 100 steps.
  • a step may e.g.
  • removing fluid from a compartment comprising removing fluid from a compartment (buffer, reaction chamber, etc.), filling fluid into a compartment, performing a stirring step, performing an annealing step, performing a cooling step, performing a drying step, collecting magnetic beads, etc.
  • the steps or at least some of the steps may be performed in serial sequence one step after the other. At least some steps may be performed timely parallel to each other, e.g. stirring and heating of a reaction chamber.
  • the number of steps may be lower than 500 steps to give an example for an upper limit.
  • the second fluidic system may not comprise a reaction chamber, preferably no reaction chamber that is configured to be heated and/or to be stirred.
  • Stirring elements may be arranged in at least one reaction chamber, e.g. an agitator element that may be moved by an external unit rotating a magnet or rotating a magnetic field using several inductive coils.
  • the at least one reaction chamber may have a volume that is less than 2 milliliter or less than e.g. 1 milliliter.
  • the volume of the at least one reaction chambers may be more than 50 microliter or more than 100 microliter to give examples for lower limits.
  • the shape of the at least one reaction chamber may be conical, e.g. in at least a third of its length. The conical portion may facilitate mixing or stirring within the reaction chamber.
  • the at least one buffer chamber may have a volume that is less than 2 milliliter or less than e.g. 1 milliliter or less than 0.5 milliliter.
  • the volume of the at least one buffer chamber may be more than 50 microliter or more than 100 microliter to give examples for lower limits.
  • the shape of the at least one buffer chamber may be essentially cylindrical, only small conical portions may be provided at the lower end of the buffer, in order to ease removal of a liquid from the respective buffer.
  • the movable unit may comprise at least one channel, preferably the first channel.
  • the first channel may comprise at least one winding, at least two windings or at least three windings.
  • other shapes may be used to implement of great volume, e.g. a meander like shape or a chamber like shape.
  • round windings may be used, e.g. to prevent formation of vortexes and/or in order to have a laminar flow at acceptable flow velocities.
  • edges may be used as well, e.g. increasing the diameter of the first channel or using lower flow velocities in order to use laminar flows and to prevent a turbulent flow.
  • turbulent flows may be used for mixing purposes within the first channel, e.g. during dissolving a dry master mix.
  • the windings may be wound up first in a first direction up to a center (fulcrum). Then the winding may be made in the opposite direction between the first windings up to the direction of an end of the first channel.
  • a large transport volume may be realized within small surface area of the movable unit/movable fluidic system.
  • the cartridge may be configured such that at least two or at least three different liquids may be stored, e.g. preliminary, within the first channel.
  • the at least two different liquids or the at least three different liquids may comprise different chemicals if compared with each other.
  • filling of the reaction chamber(s) may be performed in an efficient way, e.g. within short time.
  • the at least one winding or the other greater lumen on the movable unit may be used in combination with a closed loop from one pump port to another pump port. Thereby it may be possible to use pressure as well as suction for fluid transport. This may provide redundancy under the following circumstances, e.g. if the overall fluidic system comprises a hole or other leakage.
  • a portion of liquid may be transported that is bordered by a first portion of a gas, e.g. air on one end and by a second portion of a gas, e.g. air on the other end.
  • the stationary unit of the cartridge may be protected as a subassembly. The same technical effects as mentioned above may apply also to the stationary unit.
  • the movable unit of the cartridge may be protected as a subassembly.
  • the same technical effects as mentioned above may apply also to the movable unit.
  • the movable unit may be configured to be mounted to the stationary unit using an axial gasket that comprises at least one hole extending in the axial direction through the gasket, especially when the gasket is in the assembled position.
  • a method for preparing at least one molecular biological test such as microbiological test or biochemical test is provided, preferably using a cartridge according to any one of the embodiments mentioned above, the stationary unit or the movable unit.
  • the corresponding technical effects may also be valid for the method.
  • the molecular biological test such as microbiological test or the biochemical test may allow the detection, analysis, quantification of at least one substance such as biological substance.
  • the at least one substance may comprise or may consists of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • a substance specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), which may be associated with a particular disease or condition or with a specific disease or condition stage.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the at least one substance may be associated with infectious diseases, inflammatory diseases, sepsis, autoimmune diseases, cancer diseases (or simply cancer), or any combinations thereof.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the infectious disease may be respiratory disease such as pneumonia like hospitalized pneumonia, an implant or tissue infection, an intra-abdominal infection, or a urinary tract infection.
  • respiratory disease such as pneumonia like hospitalized pneumonia, an implant or tissue infection, an intra-abdominal infection, or a urinary tract infection.
  • the at least one substance may be indicative of a microorganism.
  • exemplary microorganisms may include but are not limited to a bacterium, virus, fungus, yeast and protozoa.
  • the at least one substance may be a pathogen.
  • the substance, specifically the substance comprising or consisting of a nucleic acid molecule may be a biomarker.
  • the at least one substance such as biological substance may be part of a fluid.
  • the at least one substance comprising or consisting of a nucleic acid molecule e.g. DNA or RNA molecule
  • the fluid may be a liquid or a gas.
  • the fluid may be a fluidic sample.
  • the fluidic sample may be a processed, nonprocessed or not yet processed fluidic sample.
  • the sample may be a biological sample (a processed, non-processed or not yet processed biological sample).
  • the biological sample may be preferably a bodily fluid (a processed, nonprocessed or not yet processed bodily fluid) or a bodily tissue (a processed, non-processed or not yet processed bodily tissue) sample.
  • the bodily tissue sample may be a liquified bodily tissue sample.
  • the biological sample may also be a cell culture medium or a cell culture supernatant.
  • the method may comprise filling (e.g. externally filling) a cartridge with at least one of, with several of or with all of the following components/chemicals:
  • a cleaning solution preferably water, more preferably filling the cleaning solution into a first storage chamber of a first fluidic system or of the first fluidic system,
  • auxiliary solution preferably a solution comprising or consisting of an alcohol, more preferably a solution comprising or consisting of isopropanol or ethanol, more preferably filling the auxiliary solution within a second storage chamber of the first fluidic system,
  • a first washing solution preferably filling the first washing solution into a third storage chamber of the first fluidic system
  • a second washing solution may be used.
  • the second washing solution may be a different solution or may be the same solution (e.g. with regard to components and/or quantity of components) compared to first washing solution, preferably filling the second washing solution into a fourth storage chamber of the first fluidic system, see e.g. US 5,234,809 (Akzo, Boom) which is included by reference herewith for all legal purposes.
  • a chemical liquid that is appropriate to perform drying of a material preferably of beads such as magnetic beads, more preferably ethanol, most preferably filling the chemical liquid into a fifth storage chamber of the first fluidic system,
  • An elution buffer (ELB), preferably filling the elution buffer into a sixth storage chamber of the first fluidic system, -
  • a master mix such as dry master mix, (e.g. comprising the ingredients as mentioned above) for performing a nucleic acid amplification or analysis reaction, preferably filling the master mix, such as dry master mix, into a seventh storage chamber of the first fluidic system,
  • a humidity sorbent material preferably comprising silica or a silica gel
  • preferably filling the humidity sorbent material e.g. powder or gel
  • the elution buffer may comprise a lightly buffered aqueous solution, e.g. in order to provide long term stability, e.g. for at least six months or for at least one year. However, water may be used if long term stability is not necessary. Tris (Tris(hydroxymethyl)aminomethane) buffered saline (TBS) may be used as an elution buffer.
  • the elution may comprise or consist of de-binding of nucleic acid molecules (NMA) from magnetizable particles, e.g. from beads.
  • NMA nucleic acid molecules
  • the master mix such as dry master mix, for performing a nucleic acid amplification or nucleic acid analysis reaction, may comprise polymerase (enzyme), transcriptase, magnesium (or another chemical comprising a bivalent cation in order to form a counter charge the charge of the DNA used e.g. in a PCR test).
  • Target specific primers and/or universal applicable primers may be added to the master mix.
  • the primers may be arranged elsewhere, e.g. within a (PCR) disc wheel comprising at least one detection chamber.
  • a method for performing at least one molecular biological test such as microbiological test or biochemical test is provided, preferably using a cartridge according to any one of the embodiments mentioned above, the stationary unit or the movable unit or a cartridge prepared according to the filling method.
  • the technical effects mentioned above may be valid.
  • the molecular biological test such as microbiological test or the biochemical test may allow the detection, analysis, quantification of at least one substance such as biological substance.
  • the at least one substance comprises or consists of a nucleic acid molecule (e.g. DNA or RNA molecule).
  • a nucleic acid molecule e.g. DNA or RNA molecule.
  • a substance specifically a substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), which is associated with a particular disease or condition or with a specific disease or condition stage.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the at least one substance is associated with infectious diseases, inflammatory diseases, sepsis, autoimmune diseases, cancer diseases (or simply cancer), or any combinations thereof.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the infectious disease is respiratory disease such as pneumonia like hospitalized pneumonia, an implant or tissue infection, an intra-abdominal infection, or a urinary tract infection.
  • the at least one substance is indicative of a microorganism.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the at least one substance can be a pathogen.
  • the substance specifically the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), is a biomarker.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • the at least one substance such as biological substance is part of a fluid.
  • the at least one substance comprises or consists of a nucleic acid molecule (e.g. DNA or RNA molecule) is part of a fluid.
  • the fluid may be a liquid or a gas.
  • the fluid may be a fluidic sample.
  • the fluidic sample may be a processed, nonprocessed or not yet processed fluidic sample.
  • the sample may be a biological sample (a processed, non-processed or not yet processed biological sample).
  • the biological sample may be preferably a bodily fluid (a processed, nonprocessed or not yet processed bodily fluid) or a bodily tissue sample (a processed, nonprocessed or not yet processed bodily fluid) sample.
  • the bodily tissue sample may be a liquified bodily tissue sample.
  • the biological sample may also be a cell culture medium or a cell culture supernatant.
  • the test method may comprise at least one, several or all of the following steps:
  • the sample may be mixed with a buffer solution or added to a (sample) buffer solution comprising or consisting of chaotropic salt(s) and/or surface-active substance(s).
  • the mixing/adding may be such that addition of alcohol may lead to binding on the magnetic beads, e.g. may fulfil the binding conditions of the substance on the beads.
  • about one third sample volume, about one third (sample) buffer solution volume and about one third alcohol volume e.g. ethanol
  • a sample preferably a cell-containing sample
  • at least one substance specifically or preferably a substance comprising or consisting of at least one nucleic acid molecule (e.g. DNA or RNA molecule)
  • heating and/or stirring e.g. in order to disrupt cells (with or without nucleus), if present, and/or to release the at least one substance, specifically a substance comprising or consisting of at least one nucleic acid molecule (e.g. DNA or RNA molecule), from the cells,
  • nucleic acid molecule binding beads preferably (diamagnetic, paramagnetic, ferro- or ferri- or antiferro-) magnetic beads or magnetizable beads, preferably paramagnetic beads with a functionalized silicon dioxide-surface in order to bind DNA selectively and/or stirring and/or heating,
  • a washing solution WS1, WS2
  • the beads preferably the (diamagnetic, paramagnetic, ferro- or ferri- or antiferro-) magnetic beads or magnetizable beads, preferably paramagnetic beads with a functionalized silicon dioxide-surface in order to bind DNA selectively and/or stirring and/or heating, e.g. in order to remove non-bound and/or unspecific material from the beads,
  • the at least one substance specifically the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule), preferably with an elution buffer solution, from the beads into the elution buffer solution, e.g. mild buffered water or distilled water depending on the requirement of long term stability or only short term stability, thereby forming an eluate,
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • an elution buffer solution e.g. mild buffered water or distilled water depending on the requirement of long term stability or only short term stability, thereby forming an eluate
  • eluate e.g. DNA or RNA molecule
  • a master mix preferably reconstituting a dry master mix with the eluate, e.g. in order to enhance and/or enable detection of the substance, specifically the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule) in the test,
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • a test including a nucleic acid amplification or nucleic acid analysis reaction, optionally including temperature cycling during the test, preferably a PCR or RT-PCR test,
  • detecting a component under test after and/or during the test preferably using fluorescence
  • Biofilters may be used to prevent pollution of the environment.
  • the cartridges may be disposable, e.g. burning of the cartridges after use may be performed in order to prevent that microorganism and/or nanoparticles (e.g. magnetic beads) enter the environment.
  • microorganism and/or nanoparticles e.g. magnetic beads
  • Figure 1 a stationary fluidic system of a stationary unit of a cartridge whereby movable unit(s) are omitted.
  • Figure 2 a start (default) switching position of a movable unit, e.g. of a switching wheel of the cartridge.
  • Figure 3 a movable fluidic system of a movable unit of the cartridge.
  • Figure 4 a further embodiment of a movable fluidic system using a continuous fluidic interface.
  • Figure 5 method steps of a method for handling chemicals, e.g. for preparing and/or performing a test, especially a molecular biological test such as microbiological test.
  • Figure 6 a (PCR) disk wheel that may be used as a second movable unit of the cartridge.
  • Figure 7 an embodiment of a gasket that may be arranged between the stationary fluidic system and the movable fluidic system in order to provide a fluid tight connection in several switching positions of the switching wheel.
  • Figure 8 an embodiment of a switching wheel rotation unit and of a (PCR) disc wheel rotation unit as well as further mechanical components of a test device.
  • Figure 9 functional blocks of an embodiment of a system for performing a test using the cartridge.
  • Figure 10 a switching position of the switching wheel that is used in order to fill the sample into a lysis chamber.
  • Figure 11 a switching position of the switching wheel that is used in order to get fluid, e.g. isopropanol, from a buffer chamber into the movable fluid system.
  • fluid e.g. isopropanol
  • Figure 12 a switching position of the switching wheel that is used in order to fill a chamber of the (PCR) disc wheel.
  • Figure 1 illustrates a first or stationary fluidic system FS1 of a stationary unit 100 of a cartridge C.
  • Movable unit(s) 200, SW and 600, DW are omitted, see figures 2, 3 and 6.
  • the cartridge C is also illustrated e.g. in figures 2 and 9, etc.
  • the cartridge C comprises sidewalls SW1 to SW4, especially of the stationary unit 100.
  • the sidewall SW 1 may be a lower sidewall in the usage position of the cartridge C.
  • the sidewall SW2 may be a left sidewall (as illustrated in figure 1), e.g. a sidewall that is inserted first into a slot SL1, see figure 9, of a machine 900 used to perform a test.
  • the sidewall SW3 may be a right sidewall, e.g. a sidewall that is inserted into the slot SL1 after the side wall SW2 has been inserted.
  • the sidewall SW4 may be an upper sidewall in the usage position of the cartridge C.
  • the cartridge C, and especially the stationary unit 100 may comprise at least two or exactly two large circular apertures 110, 112.
  • the circular aperture 110 may form a receptacle for the movable (rotatable) unit 200, SW which may comprise a second/movable fluidic system FS2.
  • the second/movable fluidic system FS2 may be movable (rotatable) relative to the first fluidic system FS1.
  • the circular aperture 112 may form a receptacle for the movable (rotatable) unit 600, DW which may comprise chambers Cl to Cl l comprising different test auxiliary materials, e.g. primers and probes, preferably labeled or coupled with at least one, at least two or at least three fluorescence material(s) per chamber Cl to Cl l, optionally combined with at least one, at least two or at least three quencher(s), e.g. using FRET (Forster resonant energy transmission), e.g. via molecular dipole coupling (especially non-radiating) or non-FRET principles.
  • the movable (rotatable) unit 600, DW may be movable (rotatable) relative to the first fluidic system FS1.
  • a rim R1 may border aperture 110. Rim R1 may be used to position a gasket 700 between the stationary unit 100 and movable (rotatable) unit 200. An embodiment of a gasket is illustrated in figure 7.
  • Rim R2 may border aperture 112. Rim R2 may be used to position a gasket between the stationary unit 100 and movable (rotatable) unit 600, e.g. a further gasket.
  • Gasket 700 may be arranged within a groove Grl arranged around aperture 110 and arranged adjacent to rim R1.
  • a further groove Gr2 is arranged around aperture 112 and arranged adjacent to rim R2 e.g. in order to hold and to position the further gasket mentioned above.
  • the first fluidic system (stationary) FS1 may comprise four groups G1 to G4 of “taps” that are arranged around aperture 110 in the following sequence and in a clockwise direction:
  • Border B is arranged between taps of the group G3 and of the group G4. A tap at the border may be omitted or may be realized as well,
  • the taps may be formed as axial portions within the stationary unit 100. Moreover, holes within gasket 700 and/or cylindrical portions of gasket 700 may be parts of the taps. Alternatively, at least one continuous fluidic interface portion may be used comprising only one continuous elongated hole instead of several taps of one of the groups G1 or G2, see e.g. description of figure 4 below.
  • the fluidic system FS1 may comprise a further channel z3 as well as channels CH10 and CH20.
  • the first fluidic system FS1 (stationary) may comprise e.g. three reaction chambers RC1 to RC3, e.g. a lysing chamber (RC1), a magnetic separation chamber (MSC, RC2) and a master mix chamber (MMC, RC3).
  • the fluidic system FS1 may comprise a plurality of buffer chambers used for storage of fluids or of dry chemicals.
  • the following buffers/chambers may be provided within cartridge C:
  • Master mix buffer chamber preferably for storage of a so called master mix MM.
  • the master mix MM may be stored as a dry powder.
  • the respective chamber may be named as a dry master mix buffer chamber DMC.
  • - Buffers (chambers) Bl to B6 e.g. used for storing liquids as explained in more detail below, e.g. aqueous solution (e.g. distilled water), isopropanol, washing solution(s), ethanol, elution buffer, etc.
  • the sample tube ST may comprise a sample S, e.g. a sample S comprising the at least one substance under test.
  • the sample tube ST may also include a solution that may enhance or allow testability of the at least one substance, e.g. a buffer solution as mentioned above, e.g. comprising chaotropic salt(s) and/or surface-active substances.
  • the sample S may be inserted in the sample tube ST, e.g. by a practitioner, e.g. using a pipette or a tweezer. Thereafter the sample tube ST may be closed, e.g. by a cap comprising a screw connection, a clip connection or another connection.
  • Other parts of the sample tube ST are described below in more detail, see e.g. figure 9 and description of figure 9.
  • a waste chamber W e.g. used to store waste liquid produced in the micro reactor that is implemented in cartridge C and that is taken into operation using machine (system, device) 900.
  • machine system, device
  • buffers that have been emptied may also be used as waste chambers during operation of the micro reactor.
  • a reservoir R e.g. in order to store a portion of the sample, especially of a lysate of the sample, for further purposes, e.g. for performing further test and/or for research or scientific purposes.
  • a chamber 120 for drying powder e.g. a powder that makes sure that the master mix may be stored as a dry powder even if cartridge C is stored in an environment having high or higher humidity.
  • An auxiliary chamber 122 of fluidic system FS1 may be used to dissolve the master mix MM during operation of the micro reactor implemented by cartridge C.
  • a flexible hose H may be connected to two fluidic ports FP1 (first) and FP2 (second).
  • the flexible hose H may be pressed and released by a peristaltic pump. Alternatively, other pumping principles may be used.
  • the channel CH10 may connect first fluidic port FP1 and the first coupling portion channel CPCH1.
  • the channel CH20 may connect second fluidic port FP2 to the second coupling portion channel CPCH2.
  • fluidic system FS1 may comprise three openings 01 to 03.
  • Waste chamber W may also comprise an opening to the environment of cartridge C.
  • Biological filters may be used in openings 01 to 03 and at the opening of waste chamber W in order to protect the environment.
  • Opening 01 may be connected to a valve VI or may be part of a valve that is operated by the machine 900 into which cartridge C is inserted to perform at least one test or a plurality of tests.
  • At least one, an arbitrary selection of or all of the following fluidic sub-systems may be provided within the first fluidic system FS1, especially arranged in the clockwise direction:
  • the first coupling portion channel CPCH1 fluidically connected to taps 1 to 8 of the first group G1 and e.g. to channel CH10, i.e. to first fluidic port FP1.
  • Channel z2 may be connected to the first coupling portion channel CPCH1 immediately below tap 1 of the first group G1 of taps.
  • Channel CH10 may be fluidically connected to the first coupling portion channel CPCH1 immediately below tap 5 of the first group G1 of taps or at another appropriate location thereby forming a passage Pl. Regardless of taps, there may be no further fluidic connection to the first coupling portion channel CPCH1.
  • the first coupling portion channel CPCH1 may comprise an intermediate volume IV, IV1 that is arranged between a passage Pl and the taps connected to or arranged on the first coupling portion channel CPCH1.
  • one void tap position arranged between the first coupling portion channel CPCH1 and the second coupling portion channel CPCH2,
  • the second coupling portion channel CPCH2 connected to taps 1 to 7 of the second group G2 and to channel CH20, i.e. to second fluidic port FP2.
  • Channel CH20 may be fluidically connected to the second coupling portion channel CPCH2 immediately below or at tap 1 of the second group G2 of taps or at another appropriate location thereby forming a passage P2. Regardless of taps, there may be no further fluidic connection to the second coupling portion channel CPCH2.
  • the second coupling portion channel CPCH2 may comprise an intermediate volume IV, IV2 that is arranged between a passage P2 and taps connected to or arranged on the second coupling portion channel CPCH2.
  • auxiliary chamber 122 may be fluidically connected to the upper portion of master mix chamber DMC.
  • the lower portion of master mix chamber DMC may be connected to channel b.
  • Channel b may be fluidically connected to tap 2 of group G3.
  • a fluidic loop may be established via channel c, a portion of the fluidic system on disc wheel DW, 600 and channel d as well as using other portions of fluidic systems FS1 and FS2.
  • - Taps 11, 12 of the third group G3 of taps e.g. fluidically connected via channels k and 1 to an upper portion of the fifth buffer B5 and to a lower portion of the fifth buffer B5 respectively
  • - Taps 13, 14 of the third group G3 of taps e.g. fluidically connected via channels m and n to an upper portion of the fourth buffer B4 and to a lower portion of the fourth buffer B4 respectively
  • border B is arranged between taps of group G3 and taps of group G4,
  • Waste buffer W may comprises an opening, preferably at its upper end as mentioned already above.
  • - Tap 4 of the fourth group G4 of taps is unused but may be used for special purposes, e.g. for a further single connection to a buffer having a “venting” opening.
  • the channel to the upper portions of a buffer or of a reaction chambers comes first, i.e. before the channel to the lower portion of the respective buffer or reaction chamber, if the switching wheel SW, 200 is rotated in a clockwise direction.
  • the channel to the lower portions of a buffer or of a reaction chambers come first, i.e. before the channel to the upper portion of the respective buffer or reaction chamber, if the switching wheel SW, 200 is rotated in a clockwise direction. Therefore, at the border B, the channel n that is connected to the lower portion of buffer B4 is adjacent to the channel o that is connected to the lower portion of buffer B3.
  • a channel z3 may fluidically connect the lower portion of sample tube holder STH, especially the lower portion of piercing pin PP, see figure 9, with a passage in reaction chamber RC1 (e.g. lysis chamber).
  • the passage in the reaction chamber RC1 may be arranged within a middle portion of the reaction chamber RC1, e.g. between the lower portion and the upper portion of the reaction chamber RC1.
  • Locking means LI, L2 e.g. flexible arms may be used to prevent rotation (moving) of the movable units 200, SW; 600, DW during transport and storage of cartridge C.
  • the locking means may interact with e.g. a respective protrusion on movable units 200, SW; 600, DW.
  • Machine 900 may be configured to unlock locking means LI, L2, e.g. by pressing down the flexible arms or in another appropriate way.
  • the cartridge C may comprise a first flat member, e.g. in the shape of a plate, e.g. a plate having a rectangular circumference or essentially rectangular, e.g. in at least 70 or at least 80 percent of its circumference. Other shapes are also possible, e.g. circular, oval, elliptical, disc like, etc.
  • the first flat member may be an outer member. A rectangular shape may allow a good orientation of the cartridge C, e.g. for a proper and/or easy insertion in a machine, e.g. machine 900.
  • the first flat member may provide a plate like support for the first fluidic system FS1 and may comprise a lateral half of the first fluidic system FS1, more specifically of the channels, e.g. a to z3 and/or of the buffers, e.g. Bl to B6 and of the reaction chambers RC1 to RC3, etc.
  • the first flat member may comprise no axial portions AP, especially not adjacent to switching wheel SW.
  • the first flat member may be made of a non-transparent material, e.g. it may be opaque, e.g. black.
  • a plastic material may be used for the first flat member e.g. using injection molding.
  • the sample tube holder STH may be the only part that is not produced using the assembly of two halves but as a single part.
  • the sample tube holder STH may be part of the first flat member.
  • the cartridge C may comprise a second flat member, preferably having the same shape as the first flat member, e.g. rectangular or having essentially the same shape. “Essentially” may mean that there are deviations only in e.g. less than 20 percent of circumference, e.g. in the region for the arrangement of the flexible hose H.
  • the second flat member may also be an outer member of the cartridge C. There may be no intermediate members of the cartridge C arranged between the first flat member and the second flat member.
  • the first flat member and the second flat member of an embodiment of the cartridge C are illustrated in figure 8. However, alternatively, one or more intermediate layer(s)/members may be arranged between the first flat member and the second flat member, e.g. in order to provide a true three-dimensional fluidic system, e.g. comprising cross overs of channels.
  • the second flat member may be plate like.
  • the second flat member may be transparent, e.g. in order to allow monitoring of the operation of the reactor and/or to allow visual inspection of the cartridge C, e.g. automatic or manual inspection.
  • the second flat member may be made of a plastic material, e.g. using injection molding.
  • Axial portions AP may be provided within the second flat member.
  • a recess may be arranged in the second flat member for arranging the sample tube holder STH of the first flat member in the assembled state of the cartridge C.
  • first flat member may be provided, e.g. as second lateral half. Deviation between the first flat member and the second flat member may be only present at the switching wheel SW or other rotatable units, e.g. test disk wheel DW (PCR disc), e.g. the taps may be only present in the second flat member.
  • test disk wheel DW PCR disc
  • Apertures 110 and 112 may extend through the first flat member and through the second flat member of the cartridge C.
  • At least one flat member may comprise or may consist of a flexible foil, e.g. the flat member may be flat on both sides, e.g. without grooves for channels and/or buffers and/or reaction chambers.
  • the cartridge C may comprise at least one or all of the following further components:
  • the movable (switching) unit e.g. switching wheel SW, 200,
  • At least one gasket e.g. gasket 700, see figure 7, preferably two gaskets, e.g. gasket 700 and the further gasket as mentioned above.
  • the switching unit/wheel SW, 200 may comprise a first flat member, e.g. in the shape of a plate, e.g. a plate having a circular, oval, elliptical, disc like shape, etc. Other shapes are possible as well, e.g. rectangular, e.g. in case of a translatable switching unit.
  • the first flat member may be an outer member of switching unit/wheel SW, 200.
  • the first flat element may provide a plate like support.
  • the first flat member of the switching unit/wheel SW, 200 may be opaque.
  • the switching unit/wheel SW, 200 may comprise a second flat member, having essentially the same shape as the first flat member.
  • the second flat member of the switching unit/wheel SW, 200 may be an outer member. There may be no intermediate members between the two outer members of switching unit/wheel SW, 200. Alternatively, one or more intermediate members may be used, e.g. in order to provide more sophisticated second fluid systems FS2.
  • the second flat member of the switching unit/wheel SW, 200 may be plate like.
  • the second flat member of the switching unit/wheel SW, 200 may be transparent, e.g. in order to allow monitoring of a process and/or inspection of the switching unit/wheel SW, 200.
  • the second flat member may comprise the same channels and/or chamber as in first flat member of the switching unit/wheel SW, 200.
  • different channels or different shaped channels may be provided in both flat members of the switching unit/wheel SW, 200.
  • Two rigid flat members may be used for the switching unit/wheel SW, 200.
  • at least one flexible foil e.g. flat on both sides, e.g. without grooves for channels and/or buffers and/or reaction chambers.
  • a gasket e.g. 700, may be arranged between the stationary part of cartridge C and the switching unit/wheel SW, 200, e.g. in order to get tight fluidic connections.
  • the gasket e.g. gasket 700, may be an axial gasket relative to a rotation axis RA of the switching unit/wheel SW, 200.
  • the (PCR) disc wheel DW, 600 is described below with reference to figure 6 in more detail.
  • the (PCR) disc wheel DW, 600 may comprise a first flat member as an outer member of disc wheel DW, 600.
  • the disc wheel DW, 600 may comprise a second flat member , having essentially the same circumferential shape as the first flat member.
  • the second flat member of the disc wheel DW, 600 may be an outer member and may be preferably a foil.
  • a gasket may be arranged between the stationary part of cartridge C and the disc wheel DW, 600 in order to get tight fluidic connections.
  • the gasket at the disc wheel DW, 600 may be an axial gasket relative to a rotation axis RA of the disc wheel DW, 600.
  • Figure 5 illustrated method steps of an exemplary process that may be performed using cartridge C. The method is described below in more detail.
  • the cartridge C may comprise:
  • a stationary unit 100 comprising e.g. a stationary fluidic system FS1, and
  • a movable unit 200 comprising e.g. a movable fluidic system FS2,
  • the movable unit 200 may be movable relative to the stationary unit 100.
  • the stationary fluidic system FS1 and the movable fluidic system FS2 may be provided to receive, to guide, and/or to retain a fluid, e.g. at least one liquid and/or at least one gas, preferably air. At least one of the stationary fluidic system FS1 and the movable fluidic system FS2 may comprises a coupling portion CPCH1, e.g. a coupling portion channel or a more complex fluidic system, of the stationary fluidic system FS1 or of the movable fluidic system FS2.
  • a coupling portion CPCH1 e.g. a coupling portion channel or a more complex fluidic system
  • the coupling portion CPCH1 may comprise a fluidic passage Pl, an intermediate volume IV, IV1 and at least two fluidic interface portions, e.g. taps of the first group Gl.
  • the cartridge C may be configured to establish a fluidic communication between the fluidic passage Pl and a lumen/channel CHI to CH5 of the fluidic system to which the coupling portion CPCH1 does not belong via a fluidic flow passing first through the fluidic passage Pl, then through the intermediate volume IV, IV 1 and thereafter through a respective one of the at least two fluidic interface portions (taps) of first group Gl taps or vice versa in the other flow direction.
  • the first fluidic system FS1 may comprise at least two fluidic compartments RC1 to RC3, Bl to B6, MBB.
  • Each of the at least two fluidic compartments RC1 to RC3, Bl to B6, MBB may be connected to at least two fluidic channels a to z of the first fluidic system FS1 in order to allow internal filling and or internal removal of chemicals from the respective fluidic compartment RC1 to RC3, Bl to B6, MBB.
  • the fluidic passage Pl of the coupling portion CPCH1 may be fluidically connected to at least one pump port FP1.
  • the pump port may be configured to be connected to a pump unit, see function block FB6 as described below with reference to figure 9.
  • the fluidic passage Pl may be fluidically connected with a first fluidic compartment of the at least two fluidic compartments RC1 to RC3, Bl to B6, MBB via a first fluidic interface portion of the at least two fluidic interface portions and via at least a first fluidic channel of the at least two fluidic channels a to z.
  • the first fluidic compartment RC1 to RC3, Bl to B6, MBB may be connected via at least a second fluidic channel of the at least two fluidic channels a to z to the pump unit FB6 in order to create a closed fluidic loop.
  • the first fluidic compartment RC1 to RC3, Bl to B6, MBB may be connected via at least a second fluidic channel of the at least two fluidic channels a to z to another outlet or inlet of the first fluidic system FS1 or of the second fluidic system FS2, preferably in order to provide the closed fluidic loop or other kind of fluid circuit.
  • the coupling portion CPCH1 may comprise an elongated channel CPCH1 that extends along a circumferential direction relative to a rotation axis RA of the movable unit 200 or that extends along a straight movement direction of the movable unit 200.
  • the elongated channel CPCH1 may comprise at least two lateral fluidic connections that are configured to be connected to the at least two fluidic interface portions, e.g. taps of the first group G1.
  • the movement may be a rotation movement.
  • a rotation angle between a first relative position RP1 of the at least two relative positions RP and a second relative position RP2 of the least one two relative positions may have a value in the range of 30 degrees to 110 degrees, in the range of 50 degrees to 100 degrees or in the range of 60 degrees to 90 degrees.
  • the movement may be again be a rotation movement.
  • the cartridge C may comprise an axial gasket 700 arranged between the stationary unit 100 and the movable unit 200.
  • the axial gasket 700 may form part of a first fluidic interface IF1 and/or of a second fluidic interface IF2 or of other fluidic interfaces, e.g. IF3, IF4 etc. as mentioned below with reference to figure 4.
  • the coupling portion CPCH1 may be a first coupling portion CPCH1.
  • the fluidic passage may be a first fluidic passage Pl.
  • the intermediate volume IV may be a first intermediate volume IV1.
  • the least two fluidic interface portions may be interface portions, e.g. taps of a first group Gl.
  • At least one of the stationary fluidic system FS1 and the movable fluidic system FS2 may comprise a second coupling portion CPCH2 of the stationary fluidic system FS1 or of the movable fluidic system FS2.
  • the second coupling portion CPCH2 may comprise a second fluidic passage P2, a second intermediate volume IV, IV2 and at least two fluidic interface portions, e.g. of a second group G2.
  • the cartridge C may be configured to establish a fluidic communication between the second fluidic passage P2 and the fluidic system to which the second coupling portion CPCH2 does not belong, e.g. a second lumen/channel CH2 of the second fluidic system FS2, via a fluidic flow passing first through the second fluidic passage P2, then through the second intermediate volume IV, IV2 and thereafter through a respective one of the at least two fluidic interface portions of the second group G2 or vice versa, depending on the flow direction.
  • the at least two relative positions RP may be relative positions of a first group of relative positions.
  • a fluidic connection may be established between the first fluidic passage, e.g. Pl, and a first portion, e.g. channel CHI, of the fluidic system, e.g. FS2, to which the first coupling portion, e.g. CPCH1, does not belong.
  • the cartridge C may be configured to establish a fluidic communication between the fluidic passage, e.g. Pl, and a second portion (e.g. lumen/channel CH2 or lumen/channel CH3) of the fluidic system, e.g. FS2, to which the coupling portion, e.g. CPCH1, does not belong via a fluidic flow passing first through the fluidic passage, e.g. Pl, then through the intermediate volume IV, e.g. IV1, and thereafter through a respective one of the at least two fluidic interface portions, e.g. of the first group Gl, or vice versa, e.g. depending on the flow direction.
  • a fluidic communication between the fluidic passage, e.g. Pl
  • a second portion e.g. lumen/channel CH2 or lumen/channel CH3 of the fluidic system, e.g. FS2, to which the coupling portion, e.g. CPCH1 does not belong via a fluidic flow passing first through the fluidic passage, e
  • the relative positions of the first group are different relative positions compared to the relative positions of the second group, e.g. there is no overlap of relative positions of the two groups of relative positions.
  • An enhanced number of degrees of freedom for performing micro reactor processes may be provided if two enlarged coupling portions or coupling systems are used and if the second fluidic system is able to provide at least two different fluidic connections via at least two different channels, e.g. via at least two of the channels CHI to CH3.
  • the first fluidic system FS1 may comprise at least one reaction chamber RC1 to RC3 or at least two reaction chambers RC1 to RC3.
  • the at least one reaction chamber RC1 to RC3 may be configured or the at least two reaction chambers RC1 to RC3 may be configured to be heated by an external heating unit, see e.g. functional block FB7 as described below with reference to figure 9.
  • the at least one reaction chamber RC 1 to RC3 may comprise a stirring element or the at least two reaction chambers RC1 to RC3 may comprise a respective stirring element that may be moved by an external stirring unit, see e.g. functional block FB8 as described below with reference to figure 9.
  • the second fluidic system 200 may not comprise a reaction chamber RC1 to RC3, especially no reaction chamber that is configured to be heated and/or to be stirred.
  • FIG. 2 illustrates a start (default) switching position SP0 of a movable unit 200, e.g. switching wheel SW of the cartridge C.
  • Switching position zero SP0 may be a start position of the switching wheel SW, 200 and/or may correspond to a switching position that is used for transport of the cartridge C.
  • Rotatable (movable) unit 200 may be rotatable around a rotation axis RA.
  • Rotation axis RA may extend perpendicular or about perpendicular to the planes in which the stationary unit 100 and/or the movable/rotatable unit 200 extend laterally.
  • the second fluidic system FS2 (rotatable, movable) may be arranged on the movable/rotatable unit 200.
  • the second fluidic system FS2 may comprise at least one channel or at least two channels, e.g. channel CHI and channel CH2 or channel CHI and channel CH3, or at least three channels, e.g. channels CHI to CH3.
  • the second fluidic system FS2 may comprise e.g. five channels CHI to CH5, see also figure 3.
  • the channel CHI may be a main channel that may be configured to store the greatest volume of fluid during rotation/movement of the second fluidic system FS2, e.g. compared to other channels of the second fluidic system FS1 if any further channels are present.
  • Channel CHI may extend through or almost through a diameter of switching wheel SW, 200.
  • the channel CH2 may extend mainly in a circumferential direction of switching wheel SW, 200, preferably bridging about one quadrant, e.g. a range of 70 degrees to 110 degrees, of 80 degrees to 100 degrees, e.g. of about 90 degrees or of 90 degrees.
  • One end of the channel CH2 may be arranged close to one end of channel CHI.
  • the other end of the channel CH2 may be arranged far from both ends of the channel CHI, e.g. at a circumferential positions that is between the positions of the two ends of the channel CHI.
  • the channel CH2 may be arranged on a first side of the channel CHI.
  • the main purpose of the channel CH2 may be to use both coupling portions CPCH1 and CPCH2 at the same time, e.g. synchronously, e.g. in order to establish a closed fluidic loop between the first fluidic system FS1 and the second fluidic system FS2 within a comparable wide switching range of e.g. at least 45 degrees or of at least 60 degrees.
  • a plurality of chambers and/or buffers may be reached via the taps of the group G3, e.g. using interfaces IF1 and IF3, see for example figure 4.
  • the channel CH3 may extend mainly in a circumferential direction of the switching wheel SW, 200, preferably bridging about one quadrant, e.g. a range of 70 degrees to 110 degrees, of 80 degrees to 100 degrees, e.g. of about 90 degrees or of 90 degrees.
  • One end of the channel CH3 may be arranged close to one end of channel CHI.
  • the other end of the channel CH3 may be arranged far from both ends of the channel CHI, e.g. at a circumferential positions that is between the positions of the two ends of the channel CHI, preferably opposite to one end of the channel CH2.
  • the channel CH3 may be arranged on a second side of the channel CHI that is opposite to the first side.
  • the main purpose of the channel CH3 may be to use both coupling portions CPCH1 and CPCH2 at the same time, e.g. synchronously, e.g. in order to establish a closed fluidic loop between the first fluidic system FS1 and the second fluidic system FS1 within a comparable wide switching range of e.g. at least 45 degrees or of at least 60 degrees.
  • channel CH3 may allow to use other switching angles compared to the switching angles that are enabled by channel CH2.
  • a plurality of chambers and/or buffers may be reached via the taps of the group G4, e.g. using the interfaces IF2 and IF4, see for example figure 11.
  • the channel CH4 may extend mainly in a circumferential direction of the switching wheel SW, 200, preferably bridging less than about one quadrant, e.g. bridging a range of 45 degrees to 85 degrees.
  • the channel CH5 may extend mainly in a circumferential direction of the switching wheel SW, 200, preferably bridging more than about one quadrant, e.g. bridging a range of 95 to 120 degrees.
  • One end of the channel CH4 and one end of the channel CH5 may be arranged closely to each other, e.g. having an offset OF2 as described below in more detail.
  • the other ends of channel CH4 and channel CH 5 may be arranged about opposite or opposite to each other.
  • Channels CH4 and CH5 may be used for special purposes within the micro reactor that is formed at least partially by cartridge C, e.g. with regard to the fluidic system, see table mentioned below with reference to figure 4.
  • One application is an aeration state.
  • a first offset OF1 may be twice as much as a second offset OF2.
  • Offset OF1 may be present between most of the taps of group G1 and between most of taps of group G2, especially between taps that are neighbors. This, may enable that the two ends of channels CH2 and CH3 that are left and right of one end of channel CHI are blocked when the end of channel CH that is arranged between these ends of channels CH2 and CH3 is arranged at a tap of group G1 or group G2.
  • the close ends of channel CHI and channel CH2 may be arranged in a distance that is equal to the small offset OF2.
  • the close ends of channel CHI and channel CH3 may be arranged in a distance that is equal to the small offset OF2.
  • the small offset OF2 may be used for taps of group G3 and G4, allowing e.g. to connect all close three ends of channels CHI to CH3 to be connected with respective taps, see e.g. figure 11.
  • the axial portion APlb of channel CHI (middle port, e.g. between axial portion AP2b of the second channel CH2 and axial portion AP3b of the third channel CH3, see e.g. figure 3) is arranged slightly besides to tap 3 of group Gl, , e.g. about 1 degree besides tap 3. This is similar to the short before 11 o’clock position of an analog clock.
  • Valve VI is in the open state. None of the channels CHI to CH5 of the second fluidic system FS1 is connected to channels of the first fluidic system FS1.
  • All buffers Bl to B6 are closed, e.g. preventing that liquid flows out of these buffers, e.g. during transport and storage of cartridge C.
  • movable (rotatable) unit 200 may comprise at least one channel, e.g. channel CHI, comprising at least one winding, at least two windings or at least three windings. Round windings may be used as illustrated in figure 2. No bridges have to be used if a distance is left between windings with flow in the same direction (e.g. clockwise or counter clock wise) for the fluid flow that flows in the opposite direction out of the center of the windings, e.g. for the back flow or counter flow. At the center, e.g. at rotation axis RA both winding portions of the channel CHI may be connected.
  • Figure 3 illustrates a movable (rotatable) fluidic system FS2 of a movable (rotatable) unit 200 of the cartridge C. More details of the channels CHI to CH5 are illustrated in order to enable more detailed description of fluidic interfaces, e.g. IF1 to IF6, etc.
  • An axial portion APla of channel CHI is arranged at the end of channel CHI that is not arranged closely between ends of channel CH2 and of channel CH3.
  • An axial portion APlb of channel CHI is arranged between the closely spaced ends of channel CH2 and of channel CH3,
  • An axial portion AP2a of channel CH2 is arranged far from axial portion APla
  • An axial portion AP2b of channel CH2 is arranged closely to axial portion APla,
  • An axial portion AP3a of channel CH3 is arranged far from axial portion APla and opposite to axial portion AP2a,
  • An axial portion AP3b of channel CH3 is arranged closely to axial portion APla,
  • An axial portion AP4a of channel CH4 is arranged closely to axial portion AP2a,
  • An axial portion AP4b of channel CH4 is arranged that is arranged closely to channel CH5,
  • An axial portion AP5a of channel CH5 is arranged closely to axial portion AP2a and opposite to axial portion AP4a,
  • An axial portion AP5b of channel CH5 is arranged closely to axial portion AP4b.
  • All axial portions AP may be arranged with the same distance to the rotation axis RA, see radius Rl.
  • other arrangements of axial portions AP are possible as well, e.g. adapting the taps of fluidic system FS2 correspondingly.
  • Other paths and/or arrangements of the channels CHI to CH5 are possible as well.
  • channel CHI may be arranged along a more rectangular or quadratic path.
  • the windings of channel CHI may be shaped meander like, etc.
  • Channel CHI may be straight.
  • a fluidic interface IF5 comprises:
  • Figure 4 illustrates a further embodiment of a movable (rotatable) fluidic system FS2b using a continuous interface portion on the side of the coupling portion, preferably on the side of an elongated coupling portion CPCHlb.
  • the continuous interface portion may be part of a fluidic interface IF1.
  • Axial portion API a of a channel CHI corresponding to channel CHI of fluidic system FS2 may form the counter interface portion of fluidic interface IF1.
  • the elongated coupling portion CPCHlb may interact with axial portion APla of channel CHI in order to provide the interface IF1 over a wider range of angles, e.g. over a range of more than 20 degrees, more than 30 degrees, etc. but less than 90 degrees to give only an example for an upper limit.
  • An angle An2 indicates a counter clockwise rotation of fluidic system FS2b. If the channel CHI is in the second position as indicated by dotted lines in figure 4, axial portion APla of channel CHI is still in fluid communication with the continuous interface portion formed by channel CPCHlb. The switching positions may be nevertheless “discrete” in order to enable fluid connections of axial portion APlb and alignment with taps of group G3 for example.
  • Channel CHI of fluidic system FS2b may be straight as illustrated or may comprise at least one winding according to a further embodiment of fluidic system FS2b.
  • At least one channel corresponding to channel CH2 and/or channel CH3 of fluidic system FS2 may be present in fluidic system FS2b.
  • Channel CH2 and/or channel CH3 of fluidic system FS2b may be straight as illustrated or may have another course, e.g. curved, according to a further embodiment of fluidic system FS2b, e.g. similar to the paths in fluid system FS2.
  • Channels corresponding to at least one of the channels CH4 and CH5 may be optionally be present or may be omitted.
  • Coupling portion channel CPCH2b may fluidically interact with channel CH2 in order to form an interface IF3 that may be used simultaneously to interface IF1.
  • the axial gasket may also comprise an elongated axial opening corresponding to the shape of coupling portion channel CPCHlb.
  • a further elongated axial opening within the axial gasket may correspond to the shape of coupling portion channel CPCH2b.
  • a bottom surface of switching wheel 200b, SWb is not illustrated but may cover the elongated axial opening(s) of the gasket outside of the region covered by the axial portion API a of channel CHI or of an axial portion AP2a of channel CH2.
  • An angle Anl is 180 degrees or may be about 180 degrees in order to position axial portion APlb opposite of axial portion APla on switching wheel 200b, SWb (rotatable unit).
  • a rotation position RP1 corresponds to the more vertical position of channel CHI.
  • a rotation position RP2 corresponds to the position of channel CH that is indicated by angle An2.
  • the movable unit 200 or 200b may comprises the first channel CHI.
  • the first channel CHI may comprise the first end portion APla and the second end portion APlb.
  • the first end portion APla may be arranged, e.g. spatially and/or fluidically closer to the at least two interface portions, e.g. taps of group Gl, see e.g. interface IF1 as illustrated in figure 4.
  • the second end portion APlb may be arranged away from the at least two interface portions, e.g. taps of group Gl, see e.g. interface IF1 as illustrated in figure 4, preferably on an opposite side of the movable unit 200 compared to a side on which the first end portion APla is arranged.
  • Channel CH5 may be used on coupling portion CPCH1 allowing e.g. “venting” of a buffer or chamber, e.g. of reaction chamber RC1 (lysis chamber). Alternatively, other chambers may be vented in this way.
  • AP5a may be arranged on tap 9 (channel i, MSC (magnetic separation chamber, RC3) of group G3,
  • AP5a may be arranged on tap 10 (channel j, MSC (magnetic separation chamber, RC3) of group G3,
  • Valve VI may be in the open state
  • the pump may be decoupled from the flexible hose H,
  • axial portion AP4b may be arranged on coupling portion CPCH2, tap 7 of group G2,
  • axial portion AP5b may be arranged on tap 13 of group G4.
  • Drying of (magnetic) beads in the MSC chamber (RC2) may be possible in this switching position SP of switching wheel SW.
  • the temperature in the MSC (RC2) may be in the range of 60 °C (degree Celsius) to 80 °C. Stirring of the beads during drying may be used optionally, e.g. in order to shorten the drying time.
  • Figure 5 illustrates method steps A to J of a method 500 for handling chemicals, e.g. for preparing and/or performing a test, especially a molecular biological test such as microbiological test.
  • Step A (preparation):
  • the cartridge C may be filled with at least one of, with several of or with all of the following components:
  • a cleaning solution preferably water within a first storage chamber Bl
  • auxiliary solution preferably alcohol, e.g. isopropanol or ethanol within the second storage chamber B2
  • a second washing solution WS2 (same or different solution compared to first washing solution) within the fourth storage chamber B4, - a chemical liquid that is appropriate to perform drying of a material, preferably of magnetic beads, e.g. ethanol within the fifth storage chamber B5,
  • elution buffer within a sixth storage chamber B6, e.g. mild buffered water or distilled water depending on the requirement of long-term stability or only short term stability,
  • MM master mix
  • MM dry master mix
  • a humidity sorbent material e.g. powder or gel
  • MiniPax may be a trademark
  • silica or a silica gel within an eighth storage chamber 120 of the first fluidic system FS1.
  • Venting of the fluidic system(s) FS1 and/or FS2 may be performed within the preparation phase.
  • pressure compensation may be made with regard to different atmospheric pressure (altitude) e.g. during production (especially filling) of the cartridge and atmospheric pressure at the test location.
  • atmospheric pressure may depend on whether conditions.
  • Valve VI may be open during venting (pressure compensation), especially switching wheel SW may be rotated a full turn in order to vent all channels a to z2 and all chamb er s/ compartments .
  • the sample containing at least one substance may be fluidically transported from the sample tube ST to the reaction chamber RC1 (lysis chamber), see description of figure 10 below.
  • the sample containing at least one substance may by lysed in the first reaction chamber (lysing chamber), preferably using heating and/or stirring.
  • the sample is a biological sample comprising cells or simply a cell sample.
  • the cells are lysed and the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule) is released from the cells.
  • Step B get an alcohol such as isopropanol or ethanol, preferably isopropanol
  • MSC magnetic separation chamber
  • an alcohol such as isopropanol or ethanol, preferably isopropanol may be used to treat the lysate, preferably using magnetic or magnetizable beads and/or stirring and/or heating.
  • the alcohol such as isopropanol or ethanol, preferably isopropanol may be transported from buffer/chamber B2 into channel CHI as is described below with reference to figure 11.
  • the beads are nucleic acid binding beads.
  • a mixture of alcohol and lysate is combined with nucleic acid binding beads.
  • the mixture of alcohol and lysate is further incubated with the beads, e.g. for a time period in the range of 10 seconds to 4 minutes or in the range of 30 seconds to 60 seconds (1 minute), e.g. in order to fix/bind the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule) to the beads.
  • the mixture of alcohol and lysate is removed from the beads.
  • the beads are magnetic beads or magnetizable beads.
  • the magnetic beads are diamagnetic, paramagnetic, ferro- or ferri- or antiferro-beads, preferably paramagnetic beads with a functionalized silicon dioxide-surface or other appropriate surface, e.g. carboxylic acid- terminated polystyrene magnetic beads (CPM), polystyrene, silica, etc. in order to bind DNA and/or RNA selectively.
  • a functionalized silicon dioxide-surface or other appropriate surface e.g. carboxylic acid- terminated polystyrene magnetic beads (CPM), polystyrene, silica, etc.
  • CPM carboxylic acid- terminated polystyrene magnetic beads
  • silica silica
  • the incubation step allows the attachment of the substance comprising or consisting of a nucleic acid molecule (e.g. DNA or RNA molecule) to the beads mentioned above.
  • Step C Washing of the beads at least once with a washing solution WS1 may be performed, preferably using stirring and/or heating, e.g. in order to remove unbound or non-specifically bound material from the beads.
  • the beads may be collected on a magnet. Washing step C may be performed in reaction chamber RC2, MSC (magnetic separation chamber), e.g. using stirring. No enhanced temperatures may be necessary for washing, e.g. room temperature may be used, e.g. about 20 degree Celsius.
  • Step D Washing of the beads using a second washing solution WS2 may be performed, preferably using stirring and/or heating, e.g. in order to remove unbound or non-specifically bound material from the beads.
  • the beads may be collected on a magnet.
  • the first washing solution WS1 may be different from the second washing solution WS2.
  • washing solutions WS1 and WS may be essentially identical to each other, e.g. comprising the same chemical and/or the same amounts (concentrations) of the chemicals.
  • Washing step C may be performed in reaction chamber RC2, MSC (magnetic separation chamber), e.g. using stirring. No enhanced temperatures may be necessary for washing, e.g. room temperature may be used, e.g. about 20 degree Celsius.
  • Step E ethanol drying:
  • the beads may be dried E, preferably using a chemical liquid, more preferably using ethanol,
  • Step F elution, de-binding, release
  • the substance which may comprises or consists of a nucleic acid molecule e.g. DNA or RNA molecule
  • an elution buffer solution F comprising mild buffered water or distilled water depending on the requirement of long term stability or only short term stability.
  • Step G (prepare master mix):
  • a master mix MM may be prepared, preferably in order to enhance and/or enable detection of a component under test.
  • the master mix MM may be prepared using the eluate, e.g. the result of step F.
  • the eluate comprising the elution buffer and the substance which comprises or consists of a nucleic acid molecule (e.g. DNA or RNA molecule) may be added to a master mix MM, preferably to a wet master mix, for performing a nucleic acid amplification or nucleic acid analysis reaction.
  • a nucleic acid molecule e.g. DNA or RNA molecule
  • a dry master mix MM which is reconstituted with the eluate comprising the elution buffer and the substance which may comprise or may consist of a nucleic acid molecule (e.g. DNA or RNA molecule) for performing a nucleic acid amplification or nucleic acid analysis reaction.
  • Dissolving of the dry master mix MM may be performed using auxiliary chamber 122 and channel CHI of the second fluidic system FS2. Thereafter, intensive mixing of the master mix solution may be performed in the third reaction chamber RC3, MMC (master mix chamber).
  • Step H (dosing of chambers in (PCR) disc wheel DW): Dosing of chambers Cl to Cl 1, see figure 6, of disc wheel DW may be performed using the second operation mode of the pump, e.g. the low volume mode. Both, the switching wheel and the disc wheel DW, may be rotated in order to transport the prepared and/or treated substance into the chambers Cl to Cl 1 or at least into some of the chambers Cl to Cl 1.
  • Step I Temporal cycling/detection:
  • a test including a nucleic acid amplification or nucleic acid analysis reaction, optionally including temperature cycling, using the eluate and optionally the master mix MM may be performed, see description of function blocks FB10 and FBI 1 as described below with reference to figure 9.
  • a heat spreader 890 may be used, preferably a rotatable heat spreader 890, see figure 8. Detection may be performed using a camera or another appropriate input device, see description of function block FB12 as described below with reference to figure 9.
  • Step J clean up: The switching wheel SW may be rotated into its default (start position), see switching position SP0 as illustrated in figure 2. The temperature elements may be switched off. The pump may be disconnected from hose H. Valve VI may be opened. Status info may be displayed to the user that the test is complete.
  • steps A to J may be performed in the alphabetical sequence, e.g. as mentioned above. However, other sequences of steps A to J may be possible as well, e.g. in order to perform other tests. Moreover, it is possible to omit at least one or several of the steps Q to J. Furthermore, other steps may be performed between steps A to J.
  • Figure 6 illustrates a (PCR) disk wheel DW, 600 that may be used as a second movable (rotatable) unit of the cartridge C.
  • the (PCR) disc wheel DW, 600 may comprise a first flat member, e.g. in the shape of a plate, e.g. a plate having a circular, oval, elliptical, disc like shape, etc. Other shapes are possible as well, e.g. rectangular, e.g. in case of a translatable switching unit.
  • the first flat member of the disc wheel DW, 600 may be an outer member of disc wheel DW, 600.
  • the first flat member may provide a plate like support.
  • the first flat member of the disc wheel DW, 600 may be opaque.
  • a first foil may be used to cover the lower side of disc wheel DW, 600.
  • a second foil may be used to cover the lower side of disc wheel DW, 600.
  • the first foil and the second foil may be transparent. Welding may be used to connect the foils fluid tight to the first flat member.
  • a gasket may be arranged between the stationary part of cartridge C and the disc wheel DW, 600 in order to get tight fluidic connections.
  • the gasket may be an axial gasket relative to a rotation axis RA of the disc wheel DW, 600.
  • DW may comprise a predetermined number of basic regions arranged in a circumferential direction, e.g. sectors of a circle.
  • the basic regions may have the same area and/or may comprise the same fluidic structures.
  • twelve regions may be provided.
  • Eleven regions may be basic regions, comprising detection chambers Cl to Cl 1 that are arranged near aperture 610 and around aperture 610.
  • Each detection chamber Cl to Cl l may be connected to two radial, e.g. straight radial channels, see e.g. radial channels CH31a, CEB lb that are connected to detection chamber Cl.
  • the radial channels may extend radially outwards from the respective chamber Cl to Cl 1 to which they are connected.
  • respective axial portions may be arranged that may extend in the axial direction (i.e. parallel to a rotation axis RA of disc wheel 600, DW), see e.g. axial portions AP3 la and AP3 lb.
  • the axial portions e.g. AP31a and AP31b may be arranged between the main parts of the respective radial channel and the cartridge C.
  • At least one fluidic interface IFDW (see figure 1) may be provided between the third fluidic system FS3 (movable/rotatable) and the first fluidic system FS1.
  • ends of channels c and d which are located near the disc wheel 600, DW and/or respective taps (axial portions) connected to these ends may form part of the at least one fluidic interface IFDW.
  • the chambers Cl to Cl 1, the radial channels and the axial portions may be part of the third fluidic system FS3 of the cartridge C.
  • the third fluidic system FS3 of disc wheel 600, DW may have a different configuration.
  • one special region of disc wheel 600, DW may be different from the other regions.
  • a calibration chamber CC and/or a “short circuit” SC may be arranged in this special regions.
  • calibration chamber CC may have a circular shape.
  • the calibration chamber CC may comprise e.g. fluorescence materials that may be used to calibrate an optical input system, e.g. a camera.
  • the special region may comprise the SC short circuit (channel) that is mentioned above.
  • the calibration chamber CC may be arranged more radially inwards compared to the short circuit (channel) SC.
  • the calibration chamber CC may be arranged at a radial position that is also used to arrange the chambers Cl to Cl l.
  • other arrangements of the calibration chamber CC and/or of the short circuit (channel) SC may be used as well.
  • the calibration chamber CC and/or the short circuit (channel) SC may be comprised within the third fluidic system FS3 which is rotatable relative to stationary unit 100 of cartridge C.
  • the circular aperture 610 may be used for coupling of a heating unit (e.g. a Peltier element) on one side of the cartridge C (e.g. side of the viewer of figure 6) to a counter part of the heating unit at the other side of the cartridge C through the aperture 610.
  • a main part of the heating unit e.g. a carrier unit, may comprise an active heating/cooling element, e.g. a Peltier element.
  • the main part may be brought into contact with the upper sides of detection chambers Cl to Cl l without using aperture 610, e.g. by contacting the free surface of disc wheel DW, 600 which surface faces to the viewer in figure 6.
  • the counterpart, e.g. heat spreader 890, see figure 3, of the heating unit may not comprise an active heating/cooling element but may comprise a good heat conductor that may conduct heat coming from the main part further to the lower sides of detection chambers Cl to Cl 1.
  • Figure 7 illustrates an embodiment of a gasket 700 that may be arranged between the stationary fluidic system FS1 and the movable (rotatable) fluidic system FS2 in order to provide a fluid tight connection in several switching positions SP of the switching wheel SW, 200.
  • Gasket 700 may have a ring shape, e.g. of a circular ring. Gasket 700 may comprise an inner circular aperture 710 that is complement to the outer ring of rim Rl. Gasket 700 may comprise an outer “castellated” edge CE. Edge CE may comprise radially extending protrusions, see e.g. protrusion Prl, and radially recessed recesses, see e.g. recess Rel, thus preventing e.g. radial movement of gasket 700 during rotation of switching wheel SW, 200.
  • the protrusions may have the same circumferential width, e.g. a width Wl.
  • the recesses may have the same circumferential width, e.g. a width W2.
  • Width Wl may be the same as width W2. Alternatively, width W2 may be larger than width Wl or may be less than width W 1.
  • the width of the protrusions may be measured in a middle axial position at the most radially outwards position of a respective protrusion.
  • the width of the recesses may be measured in a middle axial position at the most radially inwards position of a respective protrusion.
  • the sidewalls of the protrusions and/or of the recesses may extend in the axial direction of gasket 700, which corresponds to the rotation axis of switching wheel SW,200 in an assembled state of cartridge C.
  • oblique sidewalls may be used that may be slanted with regard to the axial direction, e.g. with and angle in the range of 10 degrees to 30 degrees. This may enhance tightness of gasket 700. Thus, slanting may be more than necessary to eject the second flat member of cartridge C from an injection molding machine.
  • a main circular body of gasket 700 may comprise a plurality of holes, e.g. hole Hl that are arranged at positions that correspond to the positions of taps of groups G1 to G4.
  • One protrusion may be a wider protrusion compared to the other protrusion, see wider protrusion PrW.
  • the wider protrusion PrW may ease assembling of the gasket 700, e.g. by making sure that the holes, e.g. Hl, of the gasket 700 correspond to the taps of groups G1 to G4.
  • rotatory alignment of the gasket 700 may be easy since there is only one position in which gasket 700 fits into the groove Grl.
  • cylindrical extensions Cylx (axial portions) of the gasket 700 may be configured to extend into the taps of group G1 to G4.
  • Each cylindrical extensions Cylx may comprise a central axially extending hole Hx.
  • Holes Hx may have a lower diameter, e.g. in the range of 0.7 mm (millimeter) to 0.9 mm, e.g. 0.8 mm, that is less than an upper diameter, e.g. within a range of 0.9 mm to 1.1 mm, e.g. 0.98 mm.
  • the outer diameter of cylindrical extensions Cylx may be at least factor 1.5 or at least factor 2 greater than the lower diameter of holes Hx.
  • gasket 700 the cylindrical extensions Cylx (axial portions) of gasket 700 are hold securely within the taps preventing radial movement during rotation of switching wheel SW, 200.
  • the force needed for rotation may be in the range of about 5 Nm to about 15 Nm leading to considerable friction forces on gasket 700.
  • tight fluidic connections may be provided between stationary fluidic system FS1 and rotatable fluidic system FS2.
  • gasket 700 may be essentially flat on both main sides, e.g. there may be no cylindrical extensions that extend axially from a main face of gasket 700.
  • Holes e.g. hole Hy, may be arranged to correspond to the location of taps of groups G1 to G4.
  • Holes Hx may have a lower diameter, e.g. in the range of 0.7 mm (millimeter) to 0.9 mm, e.g. 0.8 mm, that is less than an upper diameter, e.g. within a range of 0.9 mm to 1.1 mm, e.g. 0.98 mm.
  • the taps may comprise axially portions that have the same diameter (lateral width) as the holes in the gasket 700, e.g. hole Hx.
  • Gasket 700 may comprise a comparably stiff material.
  • the taps may have a diameter (lateral with) that is greater than the diameter of the holes or that is less than the diameter of the holes in the gasket 700.
  • Figure 8 illustrates an embodiment of a switching wheel rotation unit 800 and of a (PCR) disc wheel rotation unit 850 and further mechanical components of a test device 900, see figure 9.
  • Switching wheel rotation unit 800 may be part of a functional block FB4, see description of figure 9. Switching wheel rotation unit 800 may be used to drive switching wheel SW, 200 that is covered by cartridge C in figure 8. However, aperture 110 of stationary unit 100 and a toothed wheel 810 (cylindrical insert) are illustrated. Toothed wheel 810 is arranged within the aperture 110 and is in mechanical contact with the switching wheel SW, 200 e.g. arranged within a ring shaped protrusion on the bottom side of switching wheel SW.
  • Force fit and/or form fit may be used to connect both parts 200, 810 mechanically.
  • an adhesive may be used or further connection elements, etc.
  • a plurality of inner tooth, e.g. teeth 812 may be arranged at the inside of a cylinder of toothed wheel 810.
  • the number of tooth on tooth wheel 810 may be in the range of 30 to 60 tooth, e.g. 45 tooth may be used.
  • a toothed wheel TWla may be configured to interact with toothed wheel 810 in order to transmit rotation forces.
  • Figure 8 illustrates a teeth 822 of toothed wheel TWla.
  • the number of tooth of toothed wheel TWla may be equal to the number of tooth of toothed wheel 810. It may be possible to move toothed wheel TWla axially after cartridge C is inserted into machine 900. This axial movement is indicated by an arrow A8.
  • DW rotation unit 850 may be moved in the same direction at the same time.
  • mechanical interference between toothed wheel 810 and toothed wheel TWla may be provided.
  • Toothed wheel TWla may be supported by a bearing Bel, e.g. a ball bearing, a roller bearing, etc.).
  • Bearing Bel may be mounted on a support member (not illustrated), preferably on a movable support member.
  • the support member may further support a toothed wheel TWlb (not illustrated) and a motor Ml (not illustrated).
  • the motor Ml may carry the toothed wheel TWlb on a shaft.
  • Toothed wheel TWlb may be similar or may be identical to toothed wheel TW2b of DW rotation unit 850 as described below.
  • Motor Ml may be similar or may be identical to motor M2 of DW rotation unit 850 as described below.
  • a toothed belt TB1 may connect toothed wheel TWla and toothed wheel TWlb in order to transmit rotational forces from motor Ml to toothed wheel TWla.
  • a teeth 832 of toothed belt TB1 is illustrated in figure 8.
  • Other transmission units may be used as well e.g. a gear comprising gear wheels but not gear belt, etc.
  • An optional cover Col may be used to cover toothed belt TB1 and/or parts of toothed wheels TWla and toothed wheel TWlb.
  • Disc wheel DW, 600 rotation unit 850 may be part of a functional block FB5, see description of figure 9.
  • Disc wheel DW, 600 rotation unit 850 may be used to drive disc wheel DW, 600 that is covered by cartridge C in figure 8.
  • the aperture 120 of stationary unit 100 and a toothed wheel 860 (cylindrical insert) are illustrated. Toothed wheel 860 may be arranged within the aperture 120.
  • toothed wheel 860 (cylindrical insert) may be in mechanical contact with the disc wheel DW, 600 e.g. arranged within a ring shaped protrusion on the bottom side of disc wheel DW.
  • Force fit and/or form fit may be used to connect both parts 600, 860 mechanically.
  • an adhesive may be used or further connection elements, etc.
  • a plurality of inner tooth, e.g. teeth 862 may be arranged at the inside of a cylinder of toothed wheel 860.
  • the number of tooth on tooth wheel 860 may be in the range of 30 to 60 tooth, e.g. 45 tooth may be used.
  • a toothed wheel TW2a may be configured to interact with toothed wheel 860 in order to transmit rotation forces.
  • Figure 8 illustrates a teeth 872 of toothed wheel TW2a.
  • the number of tooth of toothed wheel TW2a may be equal to the number of tooth of toothed wheel 860. It may be possible to move toothed wheel TW2a axially after cartridge C is inserted into machine 900. This axial movement may be synchronously or independent of the axial movement of toothed wheel TWla, see the arrow A8. Thus, mechanical interference between toothed wheel 860 and toothed wheel TW2a may be provided.
  • Toothed wheel TW2a may be supported by a bearing Be2, e.g. a ball bearing, a roller bearing, etc.).
  • Bearing Be2 may be mounted on a support member (not illustrated), preferably on a movable support member, e.g. to the same support member as bearing Bel or to a further support member.
  • the support member that supports bearing Be2 may further support a toothed wheel TW2b and a motor M2.
  • the motor M2 may carry the toothed wheel TW2b on its shaft. Toothed wheel TW2b may be similar or may be identical to toothed wheel TWlb of SW rotation unit 800.
  • Motor M2 may be an electrical motor, especially a stepper motor.
  • a control unit for motor M2 is illustrated in figure 1, e.g. comprising power transistors that have to be cooled by cool sheets or other cooling units comprising straight slits, e.g. a passive cooling may be used.
  • a toothed belt TB2 may connect toothed wheel TW2a and toothed wheel TW2b in order to transmit rotational forces from motor M2 to toothed wheel TW2.
  • a teeth 882 of toothed belt TB2 is illustrated in figure 8.
  • An optional cover Co2 may be used to cover toothed belt TB2 and/or parts of toothed wheels TW2a and toothed wheel TW2b , e.g. in order to reduce noise emitted and/or to for other purposes.
  • a heat spreader unit 890 may be arranged within the bearing Be2. Thus, heat spreader unit 890 may be arranged stationary within toothed wheel TW2b.
  • heat spreader unit 890 is rotationally and translationally fixed to the toothed wheel TW2b it is possible to use synchronous temperature cycling for all chambers, e.g. Cl to Cl 1 of disc wheel DW, 600, e.g. in order to have a simpler assembly and/or in order to mitigate temperature isolation problems between separate cylinders of heat spreader 890, see e.g. cylinders 890a to 890e.
  • the heat spreader unit 890 may comprise discs/cylinders, e.g. a number of discs corresponds to the number of regions on disc wheel DW, 600 (in the embodiment 12 cylinders), see e.g. discs 890a to 890e of heat spreader (plus further discs, that are not illustrated).
  • the discs may have central openings allowing optical inspection of chambers CH.
  • the discs may be part of contact elements CE that are connected to a central element of the heat spreader 890 by intermediate portions of good or excellent heat conduction property.
  • the heat spreader unit 890 may be heated by at least one heating element, e.g. a Peltier element.
  • the heating element may also have cooling properties, e.g.
  • the heating element may not be part of the heat spreader 890 but may be brought into contact with the heat spreader 890, e.g. via the central element of the heat spreader 890.
  • the heat spreader unit 890 may also be combined with light guiding fibers LGF or with light guides LG, e.g. 12 light guide rods in the embodiment.
  • the number of light guide rods may correspond to the number of regions on disc wheel DW, 600, e.g. places for detection chambers Cl to Cl 1 plus one place for calibration chamber CC/short circuit SC.
  • Light guiding fibers LGF or light guides LG may be used to guide e.g. fluorescent light from chambers Cl to Cl l and/or from calibration chamber CC simultaneously or independently to an optical input device that is arranged within a space surrounded by a case of bearing Be2 or at the side of bearing Be2.
  • a camera may be used as optical input device.
  • Light may be radiated into the chambers Cl to Cl l, CC, e.g. using light guiding fibers LGF or light guides LG, e.g. simple glass cylinders or pipes in combination with e.g. at least one beam splitter (e.g. dichroic), optical filter and/or dichroic mirror(s).
  • At least one LED light emitting diode
  • An image detector may be used to generate an image.
  • the image may be evaluated to determine the intensity of light, especially of fluorescence light of the probes in the chambers of the disc wheel DW, 600, e.g. chambers Cl to Cl l. From the intensity of light the presence or absence and/or the quantity of nucleic acids in each chamber may be determined. At least two pictures or at least three pictures may be taken in timely sequence according to the respective color of the LEDs.
  • light guiding fibers LGF/ light guides LG, glass rods (cylinders) or glass bars or other appropriated light guides may be separated from the heat spreader unit 890.
  • figure 8 illustrates fill hole(s) B1F that may be used to fill the buffer Bl, preferably two small fill holes may be used, e.g. one for filling and one for venting.
  • Buffer fill hole(s) B2F may be used to fill the buffer B2, preferably two fill holes, e.g. one for filling and one for venting.
  • Buffer fill hole(s) B6F for filling of buffer the B6 are also illustrated in figure 8. Filling holes of the other buffers B3 to B5 are not illustrated in figure 8.
  • the fill holes may be closed by a foil that is glued or otherwise fastened to the fill holes.
  • a dry master mix chamber fill hole(s) DMCF may be used to fill a powder of the master mix MM into the DMC.
  • a foil may be used to close the hole, e.g. an adhesive foil.
  • Figure 9 illustrates functional blocks FB of an embodiment of a machine 900 (system) for performing a test using the cartridge C.
  • the system/ machine 900 comprises at least one, several or all of the following modules which are also named as function blocks FB in the following:
  • Cartridge feeding function block FBI the cartridge C may be processed in a perpendicular position, e.g. with side wall SW1 facing the earth. Thus, gravitation effects that liquid is hold at the bottom of the reaction chambers RC1 to RC3 and/or buffers Bl to B6.
  • the cartridge C may be insertable into a testing device, see e.g. figures 8 and 9, e.g. in order to load the testing device 900 with chemicals and/or to provide the reaction chambers for the preparation of a test and/or for the test itself .
  • the cartridge C may be removable from the testing device 900, see e.g. figures 8 and 9, e.g. in order to unload the device 900.
  • the degree or level of automation of device 900 may depend from product design and/or workflow. Manual insertion of the cartridge C is possible as well.
  • the cartridge C may be inserted and/or removed through a slot SL1, e.g. on the front side of device 900 or on another appropriate side of device 900.
  • Device 900 may be configured to process only one cartridge C per time.
  • a further assembly may be used for automatic insertion and/or removal of the cartridge C.
  • the insertion/removal assembly may not only translate the cartridge but may also perform a lateral movement of the cartridge C, e.g. in order to insert driving units (e.g. to drive the movable unit 200, switching wheel SW and/or the PCR wheel), e.g. into gears or other mechanical elements arranged on the cartridge C.
  • the driving unit may be moved relative to the cartridge in order to connect the driving unit and the cartridge C.
  • the driving unit and/or the heating unit of a/the PCR disc wheel DW, 600 may be moved from one side to the cartridge C and at least one heating element for other reaction chambers and/or buffers may be pressed from the other side against the cartridge C.
  • a reaction chamber stirring function block FB8 may be arranged on the same side as the at least one heating element.
  • Sample tube insertion and opening function block FBlb According to an embodiment of the function block FBlb, a fluidic connection between the first fluidic system FS1 and the sample tube ST may be made using a piercing pin PP, see e.g. figure 9. Alternatively, an external needle may be used.
  • the sample tube ST may be inserted manually into the cartridge C, e.g. before the cartridge C is inserted into machine (system) 900.
  • the machine (system) 900 may be configured to insert the sample tube ST into a sample tube holder STH.
  • the sample tube holder STH may be arranged on the cartridge C or may have a fluidic connection to the fluidic system of the cartridge C.
  • a sealing film of the sample tube ST may be opened during this step.
  • the opening step may be combined, e.g. with the activation step of the sample tube ST as mentioned below with regard to the function block FB3.
  • Bar code scan function block FB2 According to an embodiment of the function block FB3, a “barcode scan” of a bar code on the sample tube ST may be used to initiate the start of the process, e.g. of the test process.
  • the bar code may comprise data identifying a patient, date of the sample, etc.
  • the test(s) to be performed may be encoded within the bar codes, e.g. in order to perform a check prior to starting the machine and/or in order to enable programming of machine 900 according to the specified test.
  • the trigger for starting the machine/process to be performed may depend on the distribution of the workflow between a cockpit or user input output IF and the analyzer, e.g. the device 900.
  • Functions of the analyzer/device 900 may be separated from functions of the user interface completely or as much as possible, e.g. with regard to regulatory purposes.
  • Sample tube actuation function block FB3 Prior to each test or at the beginning of each test, a sealing foil may be perforated by at least one plunger rod in order to move a plunger Pl.
  • Plunger Pl may be arranged within the sample tube ST. Alternatively, plunger Pl may be part of machine 900.
  • plunger Pl is moved to the distal end of sample tube ST, sample liquid is expelled from sample tube ST and pressed into the first fluidic system FS1 of the cartridge C.
  • the plunger Pl is moved downwards, e.g. in the direction to the ground/earth.
  • Switching wheel rotation function block FB4 may be configured as a kind of valve apparatus or valve device.
  • a rotatable switching wheel 200, SW may be used as the movable unit 200 or may be comprised within the movable unit 200, SW.
  • a translational movable valve apparatus or valve device may be used.
  • Switching wheel 200, SW may be rotated in order to connect channels with each other, e.g. ports of fluidic connections, fluidic connections and/or reaction chambers RC1 to RC3, and/or buffer chambers Bl to B6, DMC, etc.
  • at least one, several or all of the following parameters may be used:
  • the torque for the rotation of the switching wheel 200, SW may be in the range of 5 Nm Newton meter) to 15 Nm or in the range of 7 Nm (Newton meter) to 13 Nm, preferably 10 Nm may be used.
  • Rotation in both directions may be possible, e.g. clockwise or counter clockwise,
  • Positioning accuracy may be in the range of 0.2 mm (millimeter) to 0.8 mm, e.g. a value of 0.5 mm may be used,
  • the number of different rotational positions may be in the range of 170 to 80 or in the range of 150 to 100, especially 120 positions may be used, e.g. in order to complete one turn of switching wheel SW.
  • the number of different rotation positions may be higher than the number of switching positions, e.g. twice as much or at least threefold as much, e.g. in order to enable enhanced positioning accuracy.
  • the number of switching positions may be in the range of 15 to 50 or in the range of 20 to 40.
  • the number of selectable rotation positions may be the same as the number of switching positions.
  • FIG. 8 that illustrates an embodiment of a switching wheel SW (movable/rotatable unit 200) rotation block FB4 on the right side.
  • a disc wheel 600, DW is used to provide several test chambers Cl to Cl 1 that may be filled using the switching wheel SW.
  • Each chamber e.g. Cl to Cl l, may be connected to two channels for providing a fluidic connection of the respective chamber to the first fluidic system FS1 and/or to the second fluidic system FS2.
  • Fluid from one or several of the reaction chambers RC1 to RC3 and/or from one or several of the buffer chambers Bl to B6, DMC, etc. may be used to fill at least one test chambers Cl to Cl l of the disc wheel 600, DW.
  • the chambers e.g.
  • DW may be prefilled with specific fluorescent materials/probes and/or with specific primers in order to enable a plurality of different test, e.g. three test may be performed per chamber Cl to Cl l, resulting e.g. in more than 30 different tests per cartridge C.
  • At least one, several or all of the following parameters may be used:
  • the torque for the rotation of the disc wheel 600, DW may be in the range of 5 Nm Newton meter) to 15 Nm or in the range of 7 Nm (Newton meter) to 13 Nm, preferably 10 Nm may be used.
  • Rotation of the disc wheel 600, DW in both directions may be possible, e.g. clockwise or counter clockwise,
  • DW may be in the range of 0.2 mm (millimeter) to 0.8 mm, e.g. a value of 0.5 mm may be used,
  • the number of chambers on or within disc wheel 600, DW may be in the range of 5 to 15 chambers or in the range of 7 to 13 chambers,
  • the number of different rotational positions may be in the range of 170 to 80 or in the range of 150 to 100, especially 120 positions may be used, e.g. in order to complete one turn of disc wheel 600, SW. However, a lower number of positions may be used as well, e.g. in the range 80 to 20 positions.
  • the number of different rotation positions may be higher than the number of switching positions, e.g. twice as much or at least threefold as much, e.g. in order to enable enhanced positioning accuracy.
  • the number of switching positions may be in the range of 15 to 50 or in the range of 20 to 40. Alternatively, the number of selectable rotation positions may be the same as the number of switching positions.
  • At least one lovedshort circuit“ SC may be arranged on the disc wheel 600, DW.
  • the short circuit SC may connect two channels of the first fluidic system that are used to fill at least one of the chambers Cl to Cl 1, e.g. channels c and d.
  • the “short circuit” SC may be arranged instead of one chamber, i.e. within a place on which a chamber is omitted.
  • the short circuit may allow moving liquid plugs within the switching wheel SW without rotating the SW switching wheel whereby two of the channels CHI to CH3 may be connected to channels c and d.
  • At least one calibration chamber CC may be used, preferably instead of at least one chamber, e.g. Cl to Cl l.
  • FIG. 8 illustrates an embodiment of a (PCR) disc rotation block FB5 on the left side.
  • a hose pump or peristaltic pump is used. At least one pumping wheel without a tooth, e.g. a friction wheel or at least one toothed wheel may be used in order to compress the hose and to drive a fluid flow within the hose thereby.
  • a motor may be used to drive the hose pump, e.g. peristaltic pump.
  • the pump may be operated in at least one operation state. Alternatively, the pump may be operated in at least two operation states:
  • a high pumping volume may be provided, e.g. within the range of 200 pl (micro liter) to 600 pl or within the range of 300 pl to 500 pl, e.g. a pumping volume of about 400 pl or of exactly 400 pl may be used.
  • the pumping velocity may be less than 200 pl per second or less than 100 pl per second but e.g. more than 10 pl per second to give only one example for a lower limit of the range.
  • the dosing accuracy may have a medium value, e.g. of more than 1 pl, more than 2 pl or more than 3 pl but e.g. less than 20 pl to give only one example for an upper limit.
  • a low pumping volume may be provided, e.g. within the range of 5 pl (micro liter) to 20 pl or within the range of 10 pl to 15 pl, e.g. a pumping volume of about 10 pl or of exactly 10 pl may be used.
  • the pumping velocity may be less than 20 pl per second or less than 10 pl per second but e.g. more than 1 pl per second to give only one example for a lower limit of the range.
  • the dosing accuracy may be high, i.e. have a low value, e.g. of less than 5 pl, less than 3 pl or less than 2 pl but e.g. more than 0.1 pl or more than 0.4 pl to give only one example for a lower limit.
  • a dosing accuracy of +/- 1 pl may be provided by the pump in the second operation state.
  • a further intermediate pumping volume state there may be more than two different operation states of the pump, e.g. a further intermediate pumping volume state.
  • Damage of molecules pumped by the hose pump may be lower compared to damage created by other pumps, e.g. radial pump, diagonal pump or axial pump.
  • other pumps e.g. radial pump, diagonal pump or axial pump.
  • the pump may be docked onto the hose at any time. Additionally or alternatively, the pump may be undocked from the hose at any time. Moreover, a further motor may allow undocking of the pomp from the flexible hose H and docking of the pump to the flexible hose H, e.g. in order to allow advanced operation schemes of the microreactor.
  • Reaction chamber heating function block FB7 According to an embodiment of the function block FB7, at least one reaction chamber RC1 to RC3 may be heatable and/or coolable:
  • At least two reaction chambers or at least two reaction chambers may be heated separately and independently of each other.
  • a one sided heating and/or cooling may be used. However, alternatively two sided heating and/or cooling may be used. - If a one sided cooling/heating is used, thermal isolation of the other side may allow higher heating rate and/or cooling rate,
  • Temperature accuracy +/- 5 °C or +/-2 °C or within a range of 1 °C to 5 °C, e.g. absolute value of the deviation of temperature.
  • the ramp rate of temperature may be selected appropriately.
  • function block FB8 may fulfill the following functions:
  • reaction chambers RC1 to RC3 may contain a liquid that may be stirred, e.g. using an agitator element or a follower element which follows an external magnetic field, preferably synchronously. If the reaction chamber, e.g. RC1, RC3 is not configured to contain magnetizable beads, a magnetic or magnetizable agitator element may be used. If magnetizable or magnetic beads are contained in the reaction chamber, e.g. RC2, a non-magnetic agitator that may be magnetized may be used.
  • the rotation speed of the external magnetic field and/or of the agitator element may be in the range of 50 rpm (rounds per minute) to 500 rpm, e.g. for low speed and in the range of 1500 rpm to 2500 rpm for high speed, e.g. in combination with usage of magnetizable beads.
  • the agitator element may be moved additionally to its rotation or instead of its rotation within at least one, within several or within all reaction chambers also in at least two or in all three directions of the chamber space, e.g. in x-y plane and/or in a z-direction.
  • the x-y plane may be parallel to a plane in which two plates of the cartridge abut to each other.
  • the x-direction may be an up and down direction.
  • the y-direction may be a left and right direction.
  • the z-direction may be perpendicular to the x-y plane. This may allow complete mixing of the overall volume of a reaction chamber RC1 to RC3.
  • At least two drive motors may be used to enable movement in at least two directions.
  • At least three drive motors may be used to enable movement in at least three directions.
  • An electromagnet may be used or a permanent magnet. If an electromagnet is used, the rotation speed of the magnetic field may be controlled by an electric circuit that powers at least two coils (inductors). If a permanent magnet is used, the rotation speed of the magnetic field may be controlled via the rotation speed of the permanent magnet, e.g. using a further driving motor, e.g. a fourth motor of function block FB8.
  • a further driving motor e.g. a fourth motor of function block FB8.
  • more than one driving unit may be used to create a rotating magnet field, e.g. one unit for each chamber comprising a fluid which can be stirred.
  • no or only limited movement of the driving unit may be possible, e.g. in order to move the agitator within the space of the respective reaction chamber RC1 to RC3.
  • the rotation of the external magnetic field may result in rotation, e.g. with the same speed of an internal magnet or magnetizable material within the at least one reaction chamber in which a fluid, e.g. a liquid may be stirred.
  • At least one chamber may comprise magnetizable or magnetic beads. At least two rotation states may be used in combination with a non-magnetic but magnetizable agitator element:
  • MNPs magnetic nanoparticles
  • agitator magnet
  • rotational forces e.g. due to friction with the liquid and/or centrifugal forces are not strong enough to overcome the magnetic field responsible for the attachment of the beads on the surface of the stir bar.
  • SBSE stir bar sorptive extraction
  • the magnetizable agitator element may rotate with a rotation speed in the range of 1500 rpm to 4000 rpm or within the range of 2500 rpm to 3500 rpm.
  • the MNPs may return again on the surface of magnet(s) or magnetizable agitator element thus facilitating their collection and optionally also their post extraction treatment, e.g. elution of analyte, especially of the at least one substance.
  • SBSE stir bar sorption extraction
  • DpSPE disersive micro-solid phase extraction
  • SBSDpE stir bar sorptive-dispersive micro extraction
  • a non-magnetic agitator may be used together with magnetic beads or with magnitizable beads.
  • the beads may be nanoparticles (MNP - magnetic nanoparticles having a diameter below 1 micrometer.
  • the beads may have a diameter within the range of 0,5 gm (um, micrometer) to 20 gm, e.g. within the range of 0.5 gm to 1.5 gm e.g. about lum or 1 gm.
  • the beads may have a diameter within the range of 5 gm to 15 gm, e.g. about 10 gm or 10 gm.
  • the surface area of the beads may be selected appropriately.
  • stirring principles may be used, e.g. mechanical stirring using at least one stirring element that may be mechanically and/or magnetically or in another appropriate way coupled to an external driving unit.
  • the stirring element may comprise at least one rotor blade in order to enhance stirring.
  • Liquid plug detection function block FB9 According to an embodiment of the function block FB9, at least one of the following may be valid:
  • a camera may be used to monitor filling of the disc wheel 600, DW chambers, e.g. Cl to Cl l.
  • Control of the pump e.g. hose pump (peristaltic pump) may be performed dependent on pictures taken by the camera or other appropriate electro optical device.
  • An appropriate software may be used for this purpose.
  • the pump device may be operated in its low volume mode as mentioned above, during filling of the chambers, e.g. Cl to Cl 1 of the (PCR) disc wheel 600, DW.
  • Another optical input device may be used to control filling of the chambers , e.g. Cl to Cl 1 of the (PCR) disc wheel 600, DW.
  • the liquid introduced into each chamber may be referred to as liquid plug.
  • the function of function block FB9 may be indeed a liquid plug detection.
  • function block FB10 may fulfill at least one of the following functions for at least one heating/cooling zone of a heating element used to heat at least one chamber, e.g. Cl to Cl l of (PCR) disc wheel DW:
  • At least two or three spatially separated heating zones may be used with the following operation ranges:
  • a temperature within the range of +35 °C to +68 °C may be used;
  • the temperature accuracy may e.g. be within the range of +/- 0.5 °C, especially in the range of +55 °C to +65 °C
  • a temperature within the range of +68 °C to +74 °C may be used.
  • the temperature accuracy may e.g. be within the range of +/- 0.5 °C.
  • a temperature within the range of +92 °C to +98 °C may be used.
  • the temperature accuracy may e.g. be within the range of +/- 0.5 °C.
  • a temperature cycle of a respective chamber may start with a comparably high temperature for denaturation, e.g. used for separating two DNA or RNA strings.
  • An annealing phase “annealing” with a lower temperature compared to the temperature for denaturation may follow the denaturation phase, e.g. in order to allow primers to be located at the respective positions of a single DNA or RNA strand and/or to start “copying” or completion of the protein strand/string into a two stranded DNA or RNA.
  • an elongation phase may form the third phase of one temperature cycle.
  • the temperature in the optional elongation phase may be between the temperature in the denaturation phase and in the annealing phase.
  • two step PCR may be used as well.
  • a ramp rate in the fluid in the chambers may be greater than 3 K/s (Kelvin per second) or greater than 5 K/s, e.g. within the range of 2 K/s to 20 K/s. These value/ranges may be valid in both operation states A) and B) as mentioned below with regard to function block FBI 1. Cooling may be performed using cooling rates in the range of 5 K/s to 15 K/s, e.g. of 10 K/s.
  • the temperature ramp rate of the heater or heating element may be chosen appropriately.
  • the temperature ramp rate for operation state B) mentioned below may be chosen more moderately compared to operation state A) since only one ramp up may be necessary.
  • function block FBI 1 may work according to one of the following operation principles:
  • a heat spreader unit may be used that heats or cools all chambers of the disc wheel DW synchronously, e.g. at the same time.
  • the heat spreader may comprise only one heating and/or cooling element that is used for several chambers, e.g. a Peltier element, a resistance heating element or another appropriate heating element.
  • At least one group of at least two or of at least three spatially separated heating zones may be used, preferably a sequence in at least two or at least three groups are comprised.
  • Function block FB10 and/or function block FBI 1 may have a considerable technical effect with regard to shortening of the overall processing time, e.g. due to a high repetition number of temperature cycles, e.g. during a PCR test.
  • the number of temperature cycles may be in the range of 20 cycles to 60 cycles or in the range of 30 cycles to 50 cycles. Shortening of each temperature cycle by some seconds may reduce the overall time considerably.
  • (PCR) fluorescence detection function block FB12 According to an embodiment of the function block FB12, function block FB12 may fulfill at least one of the following functions:
  • the fluorescence in each chamber of the disc wheel 600, DW may be determined for each cycle, e.g. at the end of the cycle (“annealing” phase for two step cycle/PCR or elongation phase for three step cycle/PCR).
  • the fluorescence may be determined within different wavelength ranged corresponding to different colors. Appropriate filters may be used.
  • the fluorescence may be determined at least once per cycle, e.g. at the end of the respective cycle, e.g. after each elongation phase (e.g. 72 °C-step).
  • a photo diode, photomultiplier and/or a camera chip CCD (charge coupled device) or CMOS (complementary metal oxide semiconductor) or other detection principle) or other optical input devices may be appropriate.
  • CCD charge coupled device
  • CMOS complementary metal oxide semiconductor
  • color filters may be used to select only one fluorescence dyes (material) per picture taken with the optical input device.
  • Excitation of the material in the chambers Cl to Cl 1 of the disc wheel 600, DW, e.g. chambers Cl to Cl l, may be the same for all fluorescence materials within one chamber and/or for all chambers, e.g. at the same time.
  • At least one LED (light emitting diode), e.g. at least one power LED may be used for this purpose or another appropriate light source, preferably at least two or at least three LEDs emitting different colors or at different wavelengths maxima and/or wavelength ranges.
  • Excitation using different colors may be formed in a time sequence, e.g. each after the other excitation in order to enable or to promote selective detection of occurring fluorescence
  • At least two or at least three fluorescence filters may be used in order to discriminate between at least two or at least three fluorescence materials per chamber.
  • the filters may be arranged on at least one beam splitter, e.g. a dichroic beam splitter.
  • movable filters may be used, e.g. pivotable or slidable filters that may be moved into the optical path and out of the optical path as considered necessary, e.g. automatically.
  • Function block FB12 may have a considerable technical effect with regard to shortening of the overall processing time, e.g. due to a high repetition number of cycles, e.g. during a PCR test.
  • the number of fluorescence detection cycles may be in the range of 20 cycles to 60 cycles or in the range of 30 cycles to 50 cycles. Shortening of each fluorescence cycle by some seconds may reduce the overall time considerably.
  • Control unit function block FB13
  • a central control unit CU may be used. Alternatively or additionally, distributed control functions may be used. Control unit CU may be realized using at least one processor, e.g. at least one microprocessor and/or at least one microcontroller. Input functions (keypad, touchscreen, etc.) and/or output functions (screen) may be used to e.g. set relevant parameters and/or to display relevant parameters, e.g. process status information and/or error information.
  • a non-transitory computer readable medium e.g. CD (compact disc), memory stick, other memory (random access memory (RAM), read only memory (ROM), programable ROM (PROM), erasable PROM (EPROM), electrically erasable EPROM (EEPROM), etc.
  • CD compact disc
  • RAM random access memory
  • ROM read only memory
  • PROM programable ROM
  • EPROM erasable PROM
  • EEPROM electrically erasable EPROM
  • a computer program product e.g. a program/software (also transmitted via the internet (see RFC’s (request for comments) of the IETF (Internet engineering task force) or other authority or other appropriate data transmission network), is provided and may be claimed, comprising machine readable instructions which when executed on a control unit, e.g. FBI 3 cause the control unit, e.g. FB13, to perform at least a part of or the method according to any one of the method claim and/or to control at least one of the function blocks FBI to FB12 mentioned above.
  • a control unit e.g. FBI 3
  • a system e.g. a computer, is provided and may be claimed, comprising:
  • At least one control unit e.g. FB13, and
  • a non-transitory computer readable medium (examples are mentioned above), having stored therein instructions that are executable to cause the control unit, e.g. FBI 3 to perform at least a part of or the method according to any one of the claims and/or to control at least one of the function blocks FBI to FB12 mentioned above.
  • cross marked functions or any arbitrary combination of cross marked function within one column (phase) may be controlled in the respective phase independently of each other and/or at the same time, e.g. synchronously.
  • Fig. 10 illustrates a switching position SP10 of the movable unit 200 (switching wheel SW) that is used in order to fill the sample to reaction chamber RC1 (lysis chamber).
  • the axial portion APlb of channel CHI (middle port, e.g. between axial portion AP2b of the second channel CH2 and axial portion AP3b of the third channel CH3, see e.g. figures 2 and 3) may be arranged between taps 5 and 6 of group Gl, e.g. this is similar to the short before 12 o’clock position of an analog clock).
  • Valve VI may be in the open state.
  • the end of the channel s that is distant from reaction chamber RC1 (lysis chamber) is connected via tap 6 of the fourth group G4 with the axial port AP4a of channel CH4.
  • the other axial port AP4b of channel 4 is connected to tap 2 of the first group Gl.
  • the piercing pin PP is connected with the lysis chamber (reaction chamber RC1) via channel z3,
  • the channel r is closed on tap 5 of the fourth group G4, i.e. tap 5 is not connected to any channel CHI to CH5 of switching wheel SW, 200 but is blocked by the switching wheel SW, 200.
  • the reaction chamber RC1 lysis chamber is closed at its lower end).
  • reaction chamber RC1 lysis chamber
  • tap 6 of the fourth group G4 The channel s connects the top of reaction chamber RC1 (lysis chamber) with tap 6 of the fourth group G4.
  • tap 6 of the fourth group G4 is connected via channel CH4 to first coupling portion channel CPCH1.
  • the first coupling portion channel CPCH1 is connected with channel z2 via tap 1 of the first group Gl .
  • the channel z2 is connected with opening 01 and with valve VI which is opened.
  • sample tube ST may be pressed into sample tube ST arranged in sample tube holder STH,
  • reaction chamber RC1 lysis chamber
  • reaction chamber RC1 (lysis chamber) is pressed by the sample fluid out of reaction chamber RC1 and through channel s to channel CH4, the first coupling portion channel CPCH1 and channel z2 out of cartridge C at opening 01.
  • Fig. 11 illustrates a switching position SP20 (e.g. get isopropanol) of the movable unit 200 (switching wheel SW) that is used in order to get fluid from buffer B2 (e.g. isopropanol) into the second fluid system FS2, e.g. into the fluidic system that is arranged onto movable unit 200 (switching wheel SW).
  • buffer B2 e.g. isopropanol
  • FS2 e.g. into the fluidic system that is arranged onto movable unit 200 (switching wheel SW).
  • the axial port APlb of channel CHI (middle port, e.g. between axial portion AP2b of the second channel CH2 and axial portion AP3b of the third channel CH3, see e.g. figure 2) is arranged at tap 7 of group G4 (this is similar to the short before 9 o’clock position of an analog clock).
  • Valve VI is closed.
  • the end of the channel t that is distant from the lower end of buffer B2 is connected to the axial portion APla of channel CHI.
  • the other end of channel CHI, i.e. axial portion APlb is connected with tap 4 of group G2.
  • axial portion AP3b of channel CH3 is connected with tap 8 of group G4.
  • the other axial portion AP3a of channel CH3 is connected to tap 5 of the first group G1 (first coupling portion channel CPCH1).
  • Fluidic port FP1 is connected via channel CH10, first coupling portion channel CPCH1 and channel CH3 to the channel u that leads to the upper end of buffer B2.
  • buffer B2 Lower end of buffer B2 is connected via channel t, channel CHI, second coupling portion channel CPCH2 and channel CH20 to second fluid port FP2.
  • liquid within buffer B2 e.g. isopropanol
  • channel CHI which comprises several windings.
  • the pump is stopped if the predetermined volume of liquid (e.g. isopropanol) is within channel CHI.
  • the predetermined volume of liquid e.g. isopropanol
  • Switching wheel SW may be rotated in an appropriate position to empty channel CHI and to deliver the liquid, e.g. into reaction chamber RC2 (Magnetic separation chamber) by an appropriate rotation of switching wheel SW.
  • the short circuit SC of disc wheel DW, 600 may be connected to the channels c and d, e.g. in order to provide a default position for disc wheel DW, 600.
  • Fig. 12 illustrates a switching position SP30 (e.g. PCR disc) of the movable unit 200 (switching wheel SW) that is used in order to fill fluid from channel CHlinto one of the chambers Cl to Cl 1 of the disc wheel DW, 600.
  • the switching position SP30 e.g. PCR disc
  • the axial port APlb of channel CHI (middle port, e.g. between axial portion AP2b of the second channel CH2 and axial portion AP3b of the third channel CH3, see e.g. figure 2) is arranged at tap 3 of group G3 (this is similar to the half past 4 o’clock position of an analog clock).
  • Valve VI is closed.
  • the other end of channel CHI, i.e. axial portion API a is connected with tap 3 of group G1 (first coupling portion channel CPCH1).
  • axial portion AP2b of channel CH2 is connected with tap 3 of group G3 and therefore with the end of channel c that is distant from the disc wheel DW, 600.
  • the other axial portion AP3a of channel CH2 is connected to tap 2 of the second group G2 (second coupling portion channel CPCH2).
  • Fluidic port FP1 is connected via channel CH10, first coupling portion channel CPCH1 and channel CHI to the channel d that leads to the inflow of the chamber of disc wheel DW, 600, e.g. chamber Cl.
  • the outflow of the chamber of the disc wheel e.g. of chamber Cl, is connected via channel c, channel CH2, second coupling portion channel CPCH2 and channel CH20 to second fluid port FP2.
  • liquid within channel CHI (e.g. elute) is pumped (e.g. using suction and pressure) through the other end of channel 1 into channel d to e.g. chamber Cl of disc wheel DW, 600.
  • the pump is stopped if the predetermined volume of liquid (e.g. eluate) is within channel chamber Cl.
  • (PCR) disc wheel DW, 600 may take place using the low volume mode of the pump that is connected to flexible hose H.
  • the low volume mode may allow exact filling of the chambers of disc wheel DW, 600, e.g. considering for example also a pump hysteresis.
  • eluate may be got from e.g. MMC, RC2 into channel CHI and may be pumped (e.g. using suction and pressure) into the next chamber of disc wheel DW, 600 which may have been rotated in the meantime by one chamber position. This process may be repeated until all chambers, e.g. Cl to Cl 1 of disc wheel DW, 600 are filled.
  • the PCR test may be started.
  • Both plates or other parts of the cartridge may be welded together, e.g. by laser welding using for instance linear laser welding, i.e. a linear laser beam is scanned over the connection region and a “mask” is used to activate or deactivate line segments of the linear laser beam during scanning.
  • linear laser welding i.e. a linear laser beam is scanned over the connection region and a “mask” is used to activate or deactivate line segments of the linear laser beam during scanning.
  • another appropriate technology may be used to connect both parts fluid tightly, e.g. gluing using adhesives or using at least one gasket.
  • the washing liquid L2 may be prepared by dissolving 120 g GuSCN (guanidine thiocyanate of e.g. Fluka or another company (chaotropic salt, molecule in water solution that can disrupt the hydrogen bonding network between water molecules (i.e. exerts chaotropic activity) in 100 ml L2 buffer.
  • GuSCN guanidine thiocyanate of e.g. Fluka or another company (chaotropic salt, molecule in water solution that can disrupt the hydrogen bonding network between water molecules (i.e. exerts chaotropic activity) in 100 ml L2 buffer.
  • the washing liquid L2* may be prepared by dissolving 2.45 g (gram) KI (potassium iodide from Merck) in 25 ml L2-buffer.
  • L2 buffer (0, 1 M Tris. Cl pH 6.4) may be prepared by dissolving 12.1 g TRIS (Boehringer) in 800 ml aqua bidest. (bidestillatus), adding 8.1 ml 37% (w/v) HC1 and bringing the volume to 1 liter with aqua bidest.
  • the design of the probes may use FRET (Forster resonance energy transfer) principles or non-FRET principles.
  • FRET Form resonance energy transfer
  • the probes may be destroyed by Taq DNA polymerase in the course of the PCR, separating donor and acceptor fluorophores.
  • pathogens may be subject to the analysis of at least one substance comprised therein (e.g. nucleic acid molecules such as DNA or RNA molecule):
  • HPN Hospitalized Pneumonia
  • IAI Intelligent- Abdominal Infection
  • Clostridioides difficile C. difficile
  • Clostridium perfringens
  • ITI Plant and tissue infection
  • the amplification products of the genes or genetic regions representative for the above-mentioned pathogen (bacterial) species may be detected with a radiation transfer unit (RU).
  • the detection may be carried out by via a color marking such as fluorescence marking.
  • the bacteria-specific nucleic acid molecule such as DNA or RNA molecule to be detected may be labelled with a detectable dye such as fluorescence marker/probe like TaqMan probe.
  • a TaqMan probe may consist of a fluorophore covalently attached to the 5’-end of the oligonucleotide probe and a quencher at the 3 ’-end.
  • fluorophores e.g. 6- carboxyfluorescein, acronym: FAM, or tetrachlorofluorescein, acronym: TET
  • quenchers e.g. tetramethylrhodamine, acronym: TAMRA
  • the quencher molecule quenches the fluorescence emitted by the fluorophore when excited by the cycler’s light source via Forster resonance energy transfer (FRET).
  • FRET Forster resonance energy transfer
  • TaqMan probes are designed such that they anneal within a nucleic acid such as DNA region amplified by a specific set of primers.
  • TaqMan probes can be conjugated to a minor groove binder (MGB) moiety, dihydrocyclopyrroloindole tripeptide (DPI3), in order to increase its binding affinity to the target sequence; MGB-conjugated probes have a higher melting temperature (T m ) due to increased stabilization of van der Waals forces.
  • MGB minor groove binder
  • DPI3 dihydrocyclopyrroloindole tripeptide
  • the 5' to 3' exonuclease activity of the Taq polymerase degrades the probe that has annealed to the template. Degradation of the probe releases the fluorophore from it and breaks the proximity to the quencher, thus, relieving the quenching effect and allowing fluorescence of the fluorophore.
  • fluorescence detected in the quantitative PCR thermal cycler is directly proportional to the fluorophore released and the amount of nucleic acid such as DNA template present in the PCR. This signal is then detected with the radiation transfer unit (RU).
  • a packet of assays/tests/probes may be used for a respective cartridge, e.g. for respiratory diseases.
  • the number of tests e.g. corresponding to the number of probes comprised within the cartridge
  • the number of tests may be within the range of 10 to 40 or in the range of 15 to 35 tests.
  • tests for the following viruses may be performed: Adenovirus, at least one Coronavirus (SARS CoV, MERS CoV, Cov 19, etc.), Influenza A (Orthomyxovirus), Parainfluenza (Krupps, Paramyxovirus), Respiratory Syncytial Virus (RSV) (Paramyxovirus), Rhinovirus (common cold, Picomavirus).
  • viruses or bacteria causing hospitalized pneumonia may be added, e.g. as mentioned in this application.
  • test packages may be used as well, e.g. for the detection of:

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

La présente invention concerne une cartouche (C) de manipulation d'au moins une substance, comprenant : - une unité fixe (100) comprenant un système fluidique fixe (FS1), et - une unité mobile (200) comprenant un système fluidique mobile (FS2), le système fluidique fixe (FS1) et/ou le système fluidique mobile (FS2) comprenant une partie d'accouplement (CPCH1), la partie d'accouplement (CPCH1) comprenant un passage fluidique (P1), un volume intermédiaire (IV) et au moins deux parties d'interface fluidique, dans au moins deux positions relatives (RP) différentes de l'unité mobile (200) par rapport à l'unité fixe (100), la cartouche (C) étant conçue pour établir une communication fluidique entre le passage fluidique (P1) et une lumière du système fluidique auquel la partie d'accouplement (CPCH1) n'appartient pas par l'intermédiaire d'un écoulement fluidique passant d'abord à travers le passage fluidique (P1) puis à travers le volume intermédiaire (IV) puis à travers une partie respective desdites au moins deux parties d'interface fluidique ou inversement.
PCT/EP2024/069812 2023-07-13 2024-07-12 Cartouche de manipulation d'au moins une substance, articles et procédés correspondants Pending WO2025012428A1 (fr)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5234809A (en) 1989-03-23 1993-08-10 Akzo N.V. Process for isolating nucleic acid
US20050161669A1 (en) * 2002-08-02 2005-07-28 Jovanovich Stevan B. Integrated system with modular microfluidic components
US20090194483A1 (en) * 2008-01-31 2009-08-06 Robotti Karla M Microfluidic device having monolithic separation medium and method of use
US20150190810A1 (en) * 2010-12-03 2015-07-09 Eli N. Glezer Assay Cartridge Valve System
US20170144155A1 (en) * 2014-06-05 2017-05-25 Illumina, Inc. Systems and methods including a rotary valve for at least one of sample preparation or sample analysis
US20190283024A1 (en) * 2014-05-27 2019-09-19 Illumina, Inc. Systems and methods for biochemical analysis including a base instrument and a removable cartridge

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5234809A (en) 1989-03-23 1993-08-10 Akzo N.V. Process for isolating nucleic acid
US20050161669A1 (en) * 2002-08-02 2005-07-28 Jovanovich Stevan B. Integrated system with modular microfluidic components
US20090194483A1 (en) * 2008-01-31 2009-08-06 Robotti Karla M Microfluidic device having monolithic separation medium and method of use
US20150190810A1 (en) * 2010-12-03 2015-07-09 Eli N. Glezer Assay Cartridge Valve System
US20190283024A1 (en) * 2014-05-27 2019-09-19 Illumina, Inc. Systems and methods for biochemical analysis including a base instrument and a removable cartridge
US20170144155A1 (en) * 2014-06-05 2017-05-25 Illumina, Inc. Systems and methods including a rotary valve for at least one of sample preparation or sample analysis

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"Helvetica Chimica Acta", 1995, article "A multilingual glossary of biotechnological terms: (IUPAC Recommendations"
SAMBROOK ET AL.: "Molecular Cloning: A laboratory manual", vol. 1,2,3, 15 June 2012, COLD SPRING HARBOR LABORATORY PRESS

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