[go: up one dir, main page]

WO2017198240A1 - Système pour la détection de micro-organismes photosynthétiques et non photosynthétiques dans des échantillons biologiques par photostimulation contrôlée - Google Patents

Système pour la détection de micro-organismes photosynthétiques et non photosynthétiques dans des échantillons biologiques par photostimulation contrôlée Download PDF

Info

Publication number
WO2017198240A1
WO2017198240A1 PCT/CU2017/050004 CU2017050004W WO2017198240A1 WO 2017198240 A1 WO2017198240 A1 WO 2017198240A1 CU 2017050004 W CU2017050004 W CU 2017050004W WO 2017198240 A1 WO2017198240 A1 WO 2017198240A1
Authority
WO
WIPO (PCT)
Prior art keywords
photosynthetic
stimulation
growth
detection
microorganisms
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.)
Ceased
Application number
PCT/CU2017/050004
Other languages
English (en)
Spanish (es)
Inventor
Nardo RAMÍREZ FRÓMETA
Ángel Regueiro Gómez
Carlos Abel LAMOTHE NUVIOLA
Elier Riverón Rodríguez
Carmen Yamilé Moreno Barrios
Orestes Rolando Contreras Alarcón
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.)
Centro Nacional de Investigaciones Cientificas
Original Assignee
Centro Nacional de Investigaciones Cientificas
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 Centro Nacional de Investigaciones Cientificas filed Critical Centro Nacional de Investigaciones Cientificas
Publication of WO2017198240A1 publication Critical patent/WO2017198240A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/06—Quantitative determination
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
    • G01N21/53—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke
    • G01N21/534—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke by measuring transmission alone, i.e. determining opacity
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/59—Transmissivity
    • G01N21/5907—Densitometers

Definitions

  • This invention constitutes a system (device and method) for the rapid evaluation of the state of microbiological contamination of different biological samples, based on the use of optical stimuli (430 nm ⁇ ⁇ 480 nm and 1 ⁇ ⁇ ⁇ ⁇ p ⁇ 5 ⁇ ⁇ ⁇ /), which increase the growth rate of photosynthetic and non-photosynthetic microorganisms present in a liquid culture medium facilitating their rapid detection.
  • Photo-stimulation is normally used to inhibit or contain bacterial growth (Photostimulation method and photostimulation device, CN 103212161 A; LED multiplex source and method of use of for sterilization, bioactivation and therapy, US 20050256554 A1), or to stimulate photosensitive bacterial species or with fluorescent markers.
  • Photostimulation method and photostimulation device CN 103212161 A; LED multiplex source and method of use of for sterilization, bioactivation and therapy, US 20050256554 A1
  • there is no report of devices that allow for photo-stimulation an increase
  • a light signal from an emitting LED is applied, which allows the stimulation of the growth of bacterial species, and simultaneously, through the measurement of turbidity or bioimpedance of the culture, the detection of the bacteria is possible.
  • optical stimuli can increase the activity of many species from the response of their photo-receptor proteins [Masuda, S (2013), “Light Detection and Signal Transduction in the BLUF Photoreceptors", Plant Cell Physiol. 54 (2): 171-179].
  • US20100099132A1 relates to a new method of preparing bacterial growth inducers based on the use of light, characterized in that the method comprises cultivating Hafnia spp. at a minimum and because the growth inducer is not a self-inducer.
  • the object of the invention US20130190843A1 is to provide a photo-stimulation method in which LEDs are configured to emit red or yellow light in a specific range of illuminance, in order to stimulate the synthesis of collagen in fibroblasts and promote the circulation of the blood; as well as accelerating the removal of dead cells.
  • the LEDs can be configured to emit blue light of a certain intensity to inhibit or kill P. acnes or to suppress or reduce the synthesis of melanin from melanocytes.
  • the invention WO201307491 1 A1 provides methods for light-dependent gene regulation using a photosensitive DNA-binding protein.
  • the invention provides a recombinant nucleic acid molecule comprising a sequence encoding a light sensitive DNA binding protein (LRDP) comprising: a) a LOV domain; and b) a DNA binding domain (DBD) and is operably linked to a polynucleotide.
  • LRDP light sensitive DNA binding protein
  • DBD DNA binding domain
  • the invention WO2014072934A1 describes a method for stimulating the metabolism of non-phototrophic microorganisms involved in biological processes. It claims a biological process that includes a stage of illumination with blue light of non-phototrophic microorganisms to stimulate their metabolism.
  • the main application of this invention is the treatment of municipal wastewater and in biotechnological processes for the bioproduction of molecules of interest.
  • blue light stimulation took into account the intensity of the stimulus but the influence of the waveform and the frequency in stimulation on cellular metabolism are not analyzed.
  • the effect of stimulation on samples of clinical origin with effects on rapid microbiological diagnosis is not analyzed [Tiphlova, O and Karu, T (1991), "Action of low-intensity laser radiation on Escherichia coli", Crit Rev Biomed Eng. 18 (6): 387-412].
  • the objective of the present invention is to provide a system (device and method) that allows the detection of microbial growth early in biological samples in which it is required to detect the growth of microorganisms through the use of micro-samples.
  • This system is based on the rapid detection of turbidimetric or bioimpedance changes produced by the growth of photosynthetic and non-photosynthetic microorganisms present in an optically stimulated liquid culture medium (430 nm ⁇ ⁇ 480 nm and 1 ⁇ ⁇ ⁇ ⁇ p ⁇ 5 ⁇ ⁇ ⁇ /).
  • the novelty of the present technical solution is that it presents a method for rapid diagnosis based on the combined use of optical stimulation and the measurement of turbidity or bioimpedance in biological samples, materialized from the use of a new detection system formed by the integration of hardware and software specially designed for this purpose.
  • the turbidity and / or bioimpedance changes caused by controlled optical stimulation in the culture medium are recorded in the biological sample in liquid medium, which allows the presence of bacteria to be detected, facilitating the study of their microbiological quality.
  • the proposed system is useful for the determination of the antibiogram from isolated colonies or positive samples that are obtained directly from the sources that contain them.
  • the present invention allows to obtain the antibiotic sensitivity scheme of microorganisms either from previously isolated colonies or from positive biological samples, in the latter case saving the time required for isolation and purification processes .
  • the present invention allows, from direct urine samples, to discriminate positive samples from negative ones, whether or not they are contaminated with another germ. With the system of the present invention, results based on 350 examined samples have shown 100% correspondence with the total viable cell count in CLED medium, a method conventionally used for the detection of urinary system infection.
  • the proposed system is characterized by its rapidity, since it allows the determination of urinary infection in less than an hour, and from positive samples it offers reliable results of the antibiogram in less than two hours. It is also a highly accurate system that allows corroborating the results obtained as many times as estimated. From the social point of view it is of great importance, since for those hospitalized people it allows the supply of antibiotics in a rational and timely way, avoiding prolonged hospital stays. Likewise, from the ecological point of view it has a great impact since avoiding the inappropriate use of antibiotics and at the same time limits the development of bacterial resistance.
  • SYSMEX is a reference method for the detection of bacteria and sepsis.
  • the bacteria present must be identified before starting the antibiotic supply, using various methods, from conventional biochemical tests to PCR-based DNA tests, which take an additional 3 to 24 hours.
  • the present invention describes a new measurement system (Fig. 1) that allows to optically stimulate the growth of photosynthetic and non-photosynthetic microorganisms present in a liquid culture medium, which is used as a rapid detection method that decreases the timing of Sample analysis.
  • the proposed invention comprises a device formed by an electronic card with two measurement channels: one with an array of cells for turbidimetric measurements (Fig. 2) in samples with medium or low optical density, and another with an array of cells for impedance measurement with electrodes for high optical density samples, and a common control interface ⁇ software) associated (Fig.
  • the electronic card is structured with a modular design with specific functions and objectives for each module:
  • the Central Processing and Control Unit, UCPC (1) is responsible for generating and synchronizing all control actions on the rest of the blocks.
  • the turbidity and bioimpedance measurement channels are useful for the acquisition and conditioning of the acquired variables
  • the communication module (2) establishes communication between the UCPC processor and the personal computer (3) through commands; and the power supply (4) has the function of supplying power to all devices and blocks of the design.
  • said electronic card is associated with a program tool ⁇ software) installed in a personal computer for automatic reading and storage of the measurement data via USB (communication module), which guarantees the adequate selection of parameters during the process of characterization of the detection of microorganisms in biological samples.
  • the application developed allows the programming of the stimulation parameters of the samples under study through three working windows, also guaranteeing adequate storage and review of the data acquired when requested by the user (Fig. 3).
  • the measurement system consists of a generator of the stimulus signal (5) and reference (6); in addition to a voltage-current converter (7) that supplies the controlled optical stimulus (Sine wave, 430 nm ⁇ ⁇ 480 nm, 1 ⁇ ⁇ ⁇ / ⁇ p ⁇ 5 ⁇ ⁇ ⁇ /) to an array of five LEDs (8 ) properly located in the sample holder (9), and which are responsible for transmitting the signal to stimulate the samples (10) to be analyzed.
  • the generated signal is also used as a synchronous reference for the demodulation process of the signal received from the receivers (photo detectors).
  • the optical stimulus that passes through the sample is received with the help of an opto-electronic integrated circuit (1 1), which contains a photodiode (photodetector) and a monolithic transimpedance amplifier.
  • the signal at the output of the photodetectors is selected with a multiplexer (12) and is transmitted to a conditioning section (13), passing to a synchronous demodulation block (14), filtering (15) and conversion (16) , using a hitch amplifier structure for the synchronization of the demodulation process.
  • a channel for impedance measurements from a cell has been added, using an electrode array Bipolar stainless steel (17) with a length of 10 cm and a diameter of 1 mm, and a high precision impedance converter AD5933 (18) that combines an internal frequency generator with a digital-analog converter.
  • the generator allows the biological sample to be excited with a known frequency in this case of high optical density (19).
  • the signal obtained from the analyzed sample is acquired by the AD5933 internal analog-digital converter and is processed by applying the Fourier Discrete Transform (TDF) by the internal digital processor (PDS).
  • TDF Fourier Discrete Transform
  • PDS internal digital processor
  • the applied algorithm returns a real and imaginary value of the measured impedance, which is transferred by means of a communication protocol (21) to the UCPC and from this to the personal computer (3) for graphic representation in the developed interface.
  • Another fundamental application of this invention is directed towards the determination of the sample antibiogram (susceptibility of microorganisms to antibiotics), which is achieved in a period of time less than 2 hours, using a diagnostic kit designed for these purposes.
  • the culture medium used to follow microbial growth is the modified Mueller-Hinton Broth OXOID medium, pH 7.4 ⁇ 0.2 and sterile, which additionally includes a polymer.
  • the diagnostic kit for the detection of the antibiogram of the sample to be analyzed is characterized by the use of antibiotic discs that are commercially available and that can be used in accordance with schemes that can be varied according to any need.
  • the proposed system allows to face the interferences generated by the contamination of the samples; as well as the infection caused by more than one germ.
  • the contamination interference has been resolved by adjusting the signal in magnitude and time for the detection of internationally accepted infestant levels (> 100,000 CFU / ml), promoting the exclusion of contaminated samples not infested because they generally have bacterial levels below 1,000 CFU / ml, which allows the detection of contaminated samples only when they are also infested.
  • infestant levels > 100,000 CFU / ml
  • the contaminating species attending to the Gram are saprophytes, mostly sensitive to all antibiotics, it is evident that the contaminating strains should not interfere in the detection of the resistance scheme of the infesting strains.
  • Example 1 Influence of wavelength and light intensity in the optical stimulation of bacterial growth
  • Inocula A suspension of Escherichia coli ATCC 25923 was prepared with an initial concentration of 10 4 CFU / ml in DKD culture medium.
  • the culture was carried out at 37 ° C for 18 h with the help of a Memmert incubator model INE 700.
  • the suspension of Escherichia coli was stimulated with blue wavelength of low intensity continuously, with different light intensities (1, 8; 7 and 1 1 cd respectively) and likewise with red wavelengths with intensities of 7, 1 1 and 21 cd. Turbidity growth curves of the microorganism were recorded using a time between measurements of 5 minutes. The initial concentration of the suspension was corroborated by the plate colony count. Results
  • Example 2 Influence of the frequency of the stimulus signal on the optical stimulation of bacterial growth
  • the culture was carried out at 37 ° C for 18 h with the help of a Memmert incubator model INE 700.
  • the suspension of Escher ⁇ chia coli was stimulated with a blue wavelength of low intensity continuously, with a light intensity of 1 1 cd.
  • a sweep in frequencies of the stimulus signal from 10 Hz to 10 kHz was performed.
  • Turbidity growth curves of the microorganism were recorded using a time between measurements of 5 minutes. The initial concentration of the suspension was corroborated by the plate colony count.
  • Figure 6 shows the behavior for the different stimulation frequencies analyzed. Each curve represented is the average of five trials. The frequency range that shows a higher level of stimulation within the experimental conditions evaluated was between 10 Hz and 1000 Hz. In general, statistically significant differences in the detection time were obtained in this range (approximately 40% of the average time with respect to the rest of the frequencies used).
  • Example 3 Influence of the intensity of the stimulus signal on the optical stimulation of bacterial growth
  • Inocula A suspension of Escherichia coli ATCC 25923 was prepared with an initial concentration of 10 4 CFU / ml in DKD culture medium.
  • the culture was carried out at 37 ° C for 18 h with the help of a Memmert incubator model INE 700.
  • the suspension of Escherichia coli was stimulated with a blue wavelength of low intensity continuously, with a light intensity of 1 1 cd, and a given stimulation frequency (10 Hz to 10 kHz). Under these conditions, a current scan from 2.5 mA to 20 mA (maximum stimulation current) was performed, increasing the light intensity applied to the sample.
  • Figure 7 shows the microbial behavior for the different current intensities analyzed where each curve represented is the average of 5 tests. It shows that the growth of Escherichia coli is proportional to the increase in the intensity of the stimulus. The value of the current intensity, in addition to having a significant impact on the percentage of growth stimulation, also has an impact on the duration of the latency phase, associated with a decrease in the detection time.
  • Example 4 Analysis of the growth of microorganisms with and without optical stimulation
  • Inocula Several suspensions were prepared with the different clinical strains at different initial concentrations (from 10 2 to 10 6 CFU / ml in DKD medium).
  • the culture was carried out at 37 ° C for 18 h with the help of a Memmert incubator model INE 700.
  • Figure 8 shows that for any concentration, the stimulated samples (continuous line) shorten their latency phase and therefore their detection time with respect to the samples taken as a reference without optical stimulation (curves with intermittent lines), which represents for the concentration of 10 6 CFU / ml an average of 18.75% decrease in detection time. This percentage decreases as the concentration of the sample is reduced. However, the most notable difference between both groups of curves is observed in the duration of the exponential phase of the stimulated samples; the percentage of growth stimulation being greater ( ⁇ 256%) compared to the non-stimulated samples. The results obtained (Fig.
  • Example 5 Influence of controlled optical stimulation in the detection of microorganisms in clinical urine culture samples: results of clinical studies conducted in Cuba
  • the CLED reference method standard method
  • the present system was able to detect the same number of negative samples, also in a period of 20 minutes for a 100% effectiveness rate.
  • the general correspondence of the present system with respect to the traditional CLED method was 100%.
  • E.coli To prepare the different concentrations of E.coli, the following procedure was started: Prepare a concentration of 0.5 on the McFarland scale of E.coli using the DKD culture medium and then stimulate the solution for 1 hour. (guarantees that it is in the exponential phase of growth). Then prepare the initial working concentrations (10 8 , 10 7 and 10 6 CFU / ml). To perform the growth kinetics in strips for antibiogram (ATB) 200 ⁇ of each concentration are taken. To perform the susceptibility analysis, an antibiotic disk of nalidixic acid (30 mg) plus 200 ⁇ of the desired working concentration was placed in the well of the ATB strip.
  • ATB antibiogram
  • the culture was carried out at 37 ° C for 2 h with the help of a Memmert incubator model INE 700.
  • the culture of E.coli was continuously stimulated with blue wavelength of light intensity 1 1 cd, in a frequency range of stimulation between 1 Hz and 5000 Hz.
  • Figure 10 shows the temporal record of the diffusion of several antibiotic discs (nalidixic acid, cefepime and clindamycin) in the DKD culture medium. It is observed that the detection is null (all curves overlap), that is, the diffusion of the antibiotic disk in the DKD medium; as well as the DKD medium alone, they do not introduce interference into the measurement system. This behavior guarantees that any variation detected in a growth curve through this system is due solely to the growth of the pathogen of interest.
  • Figure 1 1 shows the effect introduced by a nalidixic acid disc in the growth curve of E. coli (for 200 ⁇ of a concentration of 10 8 CFU / ml). You can see the inhibitory effect of nalidixic acid on the growth of E.coli, which becomes noticeable after 30 minutes of the start of registration, which coincides with the total diffusion of the antibiotic disc in the culture of E.coli . In the case of concentrations 10 7 and 10 6 CFU / ml the antibiotic load is sufficient to completely inhibit the growth of the microorganism during the analyzed recording time (Figs. 12 and 13). List of figures
  • Figure 1 represents the block diagram of the invention. (Drawings)
  • Figure 2 shows an image of the array of cells for turbidimetric measurements with the samples under study inside the MEMMERT incubator.
  • Figure 3 shows an image of the application developed for the configuration of the parameters of stimulation, storage and review of the data acquired from the samples under study.
  • Figure 4 represents the growth behavior of E. coli when stimulated with blue wavelength of various light intensities
  • Figure 5 depicts the effect of optical stimulation with blue wavelength and red wavelength at intensity of 1 1 cd.
  • Figure 6 represents the response of bacterial growth to different frequencies of optical stimulation with blue wavelength and intensity of 1 1 cd.
  • Figure 7 represents the influence of the stimulus signal intensity on the growth curve of E. coli when stimulated with blue wavelength.
  • Figure 8 shows the growth curves of E. coli obtained during the study of the photostimulation effect with blue wavelength for different concentrations (from 1 0 2 to 10 6 CFU / ml).
  • Figure 9 shows the response time of several microorganisms to optical stimulation with blue wavelength.
  • Figure 10 represents the temporal record of the diffusion of several antibiotic discs (nalidixic acid, cefepime and clindamycin) in the DKD culture medium with the use of optical stimulation.
  • Figure 1 1 shows the effect introduced by a nalidixic acid disc in the growth curve of E. coli (for 200 ⁇ of an initial concentration of 10 8 CFU / ml) with the use of optical stimulation.
  • Figure 12 shows the effect introduced by a nalidixic acid disc in the growth curve of E. coli (for 200 ⁇ of an initial concentration of 10 7 CFU / ml) with the use of optical stimulation.
  • Figure 13 shows the effect introduced by a nalidixic acid disc in the growth curve of E. coli (for 200 ⁇ of an initial concentration of 10 6 CFU / ml) with the use of optical stimulation
  • Figure 2 Image of the array of cells for turbidimetric measurements with the samples under study inside a MEMMERT incubator.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biotechnology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Toxicology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Sustainable Development (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne le domaine de la microbiologie et de l'hygiène des aliments, elle constitue un système pour l'évaluation rapide de l'état de contamination microbiologique de différents échantillons biologiques à partir de l'utilisation combinée de la photostimulation-détection des micro-organismes (photosynthétiques et non photosynthétiques) présents dans un milieu de culture liquide. Dans le cas de la photostimulation (430 nm < λ < 480 nm et 1 μW< p < 5 μW) d'un échantillon biologique dans un milieu liquide, les changements de turbidité et/ou de bioimpédance causés dans le milieu de culture par cette croissance sont enregistrés, ce qui permet de détecter de manière rapide la présence de bactéries dans des échantillons biologiques à partir de l'analyse de la culture, ce qui facilite l'évaluation de la qualité microbiologique de ces échantillons. En outre, l'invention est utile pour la détermination de l'antibiogramme à partir de colonies isolées ou d'échantillons positifs qui sont obtenus directement à partir des sources qui les contiennent.
PCT/CU2017/050004 2016-05-19 2017-05-18 Système pour la détection de micro-organismes photosynthétiques et non photosynthétiques dans des échantillons biologiques par photostimulation contrôlée Ceased WO2017198240A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CU2016-0070 2016-05-19
CU2016000070A CU24452B1 (es) 2016-05-19 2017-05-18 Sistema electrónico para la detección de microorganismos en muestras biológicas por fotoestimulación controlada

Publications (1)

Publication Number Publication Date
WO2017198240A1 true WO2017198240A1 (fr) 2017-11-23

Family

ID=59284951

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CU2017/050004 Ceased WO2017198240A1 (fr) 2016-05-19 2017-05-18 Système pour la détection de micro-organismes photosynthétiques et non photosynthétiques dans des échantillons biologiques par photostimulation contrôlée

Country Status (2)

Country Link
CU (1) CU24452B1 (fr)
WO (1) WO2017198240A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4089179A1 (fr) 2021-05-12 2022-11-16 Instituto Politécnico De Leiria Procédés de détection microbiologique et de détermination de la sensibilité antimicrobienne dans des échantillons cliniques, environnementaux ou alimentaires

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040009572A1 (en) * 2002-03-11 2004-01-15 Carmelo Jose Felice Apparatus for the analysis of microorganisms growth and procedure for the quantification of microorganisms concentration
US20050256554A1 (en) 2003-12-23 2005-11-17 American Environmental Systems, Inc LED multiplex source and method of use of for sterilization, bioactivation and therapy
WO2007070452A1 (fr) * 2005-12-09 2007-06-21 Bionavitas, Inc. Systemes, dispositifs et procedes de production de biomasse
US20100099132A1 (en) 2007-01-26 2010-04-22 University Of Leicester Bacterial growth inducer
WO2010097687A1 (fr) * 2009-02-25 2010-09-02 Alifax Holding Spa Procédé de recherche de bactéries dans un échantillon biologique et dispositif associé
WO2010132955A1 (fr) * 2009-05-21 2010-11-25 Omega 3 Innovations Pty Ltd Appareil, système et procédé de photosynthèse
WO2012101459A2 (fr) * 2011-01-28 2012-08-02 Algaecytes Limited Procédé pour la production de microalgues, cyanobactéries et de métabolites de ceux-ci
WO2013074911A1 (fr) 2011-11-18 2013-05-23 Board Of Regents, The University Of Texas System Système pour l'expression génique, inductible par la lumière bleue
CN103212161A (zh) 2012-01-19 2013-07-24 福华电子股份有限公司 光刺激方法及光刺激装置
US20130190843A1 (en) 2012-01-19 2013-07-25 Forward Electronics Co., Ltd. Photo-stimulation method and device
WO2014072934A1 (fr) 2012-11-07 2014-05-15 Girinsky Olivier Utilisation de lumiere bleue pour la stimulation du metabolisme de microorganismes non phototrophes

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040009572A1 (en) * 2002-03-11 2004-01-15 Carmelo Jose Felice Apparatus for the analysis of microorganisms growth and procedure for the quantification of microorganisms concentration
US20050256554A1 (en) 2003-12-23 2005-11-17 American Environmental Systems, Inc LED multiplex source and method of use of for sterilization, bioactivation and therapy
WO2007070452A1 (fr) * 2005-12-09 2007-06-21 Bionavitas, Inc. Systemes, dispositifs et procedes de production de biomasse
US20100099132A1 (en) 2007-01-26 2010-04-22 University Of Leicester Bacterial growth inducer
WO2010097687A1 (fr) * 2009-02-25 2010-09-02 Alifax Holding Spa Procédé de recherche de bactéries dans un échantillon biologique et dispositif associé
WO2010132955A1 (fr) * 2009-05-21 2010-11-25 Omega 3 Innovations Pty Ltd Appareil, système et procédé de photosynthèse
WO2012101459A2 (fr) * 2011-01-28 2012-08-02 Algaecytes Limited Procédé pour la production de microalgues, cyanobactéries et de métabolites de ceux-ci
WO2013074911A1 (fr) 2011-11-18 2013-05-23 Board Of Regents, The University Of Texas System Système pour l'expression génique, inductible par la lumière bleue
CN103212161A (zh) 2012-01-19 2013-07-24 福华电子股份有限公司 光刺激方法及光刺激装置
US20130190843A1 (en) 2012-01-19 2013-07-25 Forward Electronics Co., Ltd. Photo-stimulation method and device
WO2014072934A1 (fr) 2012-11-07 2014-05-15 Girinsky Olivier Utilisation de lumiere bleue pour la stimulation du metabolisme de microorganismes non phototrophes

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
BRAATSCH, S: "Responses of the Rhodobacter sphaeroides Transcriptome to Blue Light under Semiaerobic Conditions", J. BACTERIOL, vol. 186, no. 22, 2004, pages 7726 - 7735
HORST, M.: "Photoreceptor proteins star actors of modern times: a review of the functional dynamics in the structure of representative members of six different photoreceptor families", ACC. CHEM. RES, vol. 37, no. 1, 2004, pages 13 - 20
JUNG, A: "Structure of a bacterial BLUF photoreceptor: Insights into blue light-mediated signal transduction", PROC NATL ACAD SCI U S A, vol. 102, no. 35, 2005, pages 12350 - 12355
KARU, T: "Primary and Secondary Mechanisms of Action of Visible to Near-IR Radiation on Cells", JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B: BIOLOGY, vol. 49, no. 1, 1999, pages 1 - 17, XP028735364, DOI: doi:10.1016/S1011-1344(98)00219-X
KUSHIBIKI, T: "Chondrogenic mRNA expression in prechondrogenic cells after blue laser irradiation", J PHOTOCHEM PHOTOBIOL B., vol. 98, no. 3, 2010, pages 211 - 215, XP026943744, DOI: doi:10.1016/j.jphotobiol.2010.01.008
MASUDA, S: "AppA is a blue light photoreceptor that antirepresses photosynthesis gene expression in Rhodobacter sphaeroides", CELL, vol. 110, no. 5, 2002, pages 613 - 623
MASUDA, S: "Light Detection and Signal Transduction in the BLUF Photoreceptors", PLANT CELL PHYSIOL., vol. 54, no. 2, 2013, pages 171 - 179
O TIPHLOVA ET AL: "Action of low-intensity laser radiation on Escherichia coli.", CRITICAL REVIEWS IN BIOMEDICAL ENGINEERING., vol. 18, no. 6, 1 January 1991 (1991-01-01), pages 387 - 412, XP055405725 *
PEREIRA, M: "Influence of 670 nm Low-Level Laser Therapy on Mast Cells and Vascular Response of Cutaneous Injuries", JOURNAL OF PHOTOCHEM. & PHOTOBIOLOGY B: BIOLOGY, vol. 98, no. 3, 2010, pages 188 - 192, XP026943740, DOI: doi:10.1016/j.jphotobiol.2009.12.005
TIPHLOVA, O; KARU, T: "Action of low-intensity laser radiation on Escherichia coli", CRIT REV BIOMED ENG., vol. 18, no. 6, 1991, pages 387 - 412

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4089179A1 (fr) 2021-05-12 2022-11-16 Instituto Politécnico De Leiria Procédés de détection microbiologique et de détermination de la sensibilité antimicrobienne dans des échantillons cliniques, environnementaux ou alimentaires

Also Published As

Publication number Publication date
CU20160070A7 (es) 2017-12-08
CU24452B1 (es) 2019-11-04

Similar Documents

Publication Publication Date Title
Volk Screening of microalgal culture media for the presence of algicidal compounds and isolation and identification of two bioactive metabolites, excreted by the cyanobacteria Nostoc insulare and Nodularia harveyana
US10995357B2 (en) Rapid antimicrobial susceptibility testing based on a unique spectral intensity ratio analysis via single fluorescence membrane dye staining and flow cytometry
US10995358B2 (en) Rapid antibiotic susceptibility test using membrane fluorescence staining and spectral intensity ratio improved by flow cytometry dead to live population ratio
WO2017198240A1 (fr) Système pour la détection de micro-organismes photosynthétiques et non photosynthétiques dans des échantillons biologiques par photostimulation contrôlée
JP6706267B2 (ja) 迅速内部蛍光法を使用した耐性遺伝子を含む細菌の無試薬同定
CN111088318A (zh) 利用ttc还原反应检测细菌药物敏感性的方法
RU2393229C1 (ru) Способ определения адгезии staphylococcus spp. к гемопротеинам
Abdelraheem et al. Phenotypic and genotypic detection of biofilm formation and methicillin resistance among Staphylococcus aureus isolates
Jones et al. Further evaluation of the test for detection of antibody-coated bacteria in urine sediment
CA3142867A1 (fr) Microscopie pour un test rapide de sensibilite aux antibiotiques utilisant une coloration par fluorescence de membrane et un rapport d&#39;intensite spectrale
CN113466309B (zh) 用于病原体检测的方法
US20100330601A1 (en) Control system for uv lamps, and check system for determining the viability of microorganisms
Jabbar et al. Antibiotics resistance profile of bacterial strains produced of biofilm isolated from patients in Al-Diwaniya city, Iraq
ES2818374T3 (es) Método de neutralización de antibióticos en un medio de cultivo
RU2603100C1 (ru) Способ оценки эффективности антимикробного воздействия антисептиков на бактерии, существующие в форме биопленки
Tiposoth et al. Rapid and quantitative fluorescence detection of pathogenic spore-forming bacteria using a xanthene-Zn (II) complex chemosensor
RU2844977C1 (ru) Способ определения Esherichia Coli и колиформных бактерий на отечественной селективной хромогенной питательной среде &#34;Хромагар&#34;
Purkayastha et al. Cultural and biochemical characterization of sheep Escherichia coli isolated from in and around Bau campus
Younus et al. Identification and Antibiotic Sensitivity of Staphylococcus aureus Isolated from Burned-Wounded Patients
RU2804102C1 (ru) Способ определения чувствительности микроорганизмов к антибиотикам при лечении гнойно-воспалительных заболеваний животных
Jahan et al. Isolation, identification and antibiogram study of bacteria from diarrhoeic goat at selected areas of Sylhet.
ES2411954T3 (es) Detección de microorganismos con un dispositivo basado en fluorescencia
RU2550254C1 (ru) СПОСОБ ОПРЕДЕЛЕНИЯ ЧУВСТВИТЕЛЬНОСТИ ШТАММОВ Pseudomonas aeruginosa К АНТИБИОТИКАМ
Mustafa et al. Application of Silver Oxide Nanoparticles as Antibiofilm and Detection of Beta Lactamase Gene in Escherichia Coli Isolated from Diabetic Mellitus Patients
Sun et al. Research Academy for Young Scientists

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17735372

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 17735372

Country of ref document: EP

Kind code of ref document: A1