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WO2025050373A1 - Module de réaction, système de préparation d'échantillon, procédé de préparation d'échantillon et système d'analyse de substance biochimique - Google Patents

Module de réaction, système de préparation d'échantillon, procédé de préparation d'échantillon et système d'analyse de substance biochimique Download PDF

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Publication number
WO2025050373A1
WO2025050373A1 PCT/CN2023/117644 CN2023117644W WO2025050373A1 WO 2025050373 A1 WO2025050373 A1 WO 2025050373A1 CN 2023117644 W CN2023117644 W CN 2023117644W WO 2025050373 A1 WO2025050373 A1 WO 2025050373A1
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WO
WIPO (PCT)
Prior art keywords
fluid
reaction
module
sample preparation
channel
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/CN2023/117644
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English (en)
Chinese (zh)
Inventor
杨谌
李桥
崔金凤
周茂顺
李川
龙小娟
龚梅花
卢剑
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MGI Tech Co Ltd
Original Assignee
MGI Tech Co Ltd
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 MGI Tech Co Ltd filed Critical MGI Tech Co Ltd
Priority to PCT/CN2023/117644 priority Critical patent/WO2025050373A1/fr
Publication of WO2025050373A1 publication Critical patent/WO2025050373A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C12M1/38—Temperature-responsive control

Definitions

  • the present application relates to the technical field of biochemical substance analysis, and in particular to a reaction module, a sample preparation system, a sample preparation method and a biochemical substance analysis system.
  • reaction module In order to solve the above deficiencies of the prior art, it is necessary to provide a reaction module, and an automated, integrated and miniaturized sample preparation system and sample preparation method using the reaction module.
  • the embodiment of the present application also provides a biochemical substance analysis system using the sample preparation system.
  • an embodiment of the present application provides a reaction module, wherein the reaction module includes a fluid reaction structure, and a reaction channel is provided on the fluid reaction structure.
  • the reaction channel is bent and arranged on the fluid reaction structure.
  • the fluid reaction structure includes a substrate and a groove formed on the substrate, a hollow reaction pipeline is provided in the groove, and the hollow portion of the reaction pipeline constitutes the reaction channel.
  • the depth of the groove is greater than or equal to the outer diameter of the reaction pipeline.
  • the reaction pipeline is detachably disposed in the groove.
  • the fluid reaction structure includes a stacked substrate and a cover plate, a groove is formed on the substrate, and a surface of the cover plate close to the substrate is sealed and connected to the substrate so that the groove forms the reaction channel;
  • the fluid reaction structure includes a stacked substrate and a cover plate, and two support walls located between the substrate and the cover plate, and the opposite end surfaces of each support wall are respectively connected to the substrate and the cover plate, and the substrate, the two support walls and the cover plate together form the reaction channel.
  • the reaction channel along the extension direction of the reaction channel, includes a plurality of reaction areas, and temperatures of at least two of the reaction areas are different.
  • the reaction module further includes a temperature control component, and the temperature control component is used to control the temperature of the fluid reaction structure to regulate the temperature in the reaction channel.
  • an embodiment of the present application provides a sample preparation system, comprising: a fluid storage module, a fluid distribution module, a fluid driving module and a reaction module, wherein the fluid storage module is used to store fluid, the reaction module has a reaction channel, and the reaction channel is respectively connected to the fluid distribution module and the fluid driving module; the fluid driving module is used to create a pressure gradient so that the fluid in the fluid storage module is transported to the reaction channel via the fluid distribution module; the reaction module is used to perform a biochemical reaction of the fluid transported into the reaction channel.
  • the fluid driving module is also used to create a pressure gradient so that the fluid after the reaction in the reaction channel is discharged into the container of the fluid storage module that stores another fluid through the fluid distribution module; the fluid storage module is also used to mix the fluid after the reaction and the other fluid to obtain a mixed fluid.
  • the fluid storage module includes a fluid storage device and a fluid switching device
  • the fluid storage device includes a plurality of containers for accommodating the fluid
  • the fluid switching device is used to adjust the positions of the plurality of containers so that one of the plurality of containers is aligned with the fluid distribution module
  • the fluid switching device is also used to mix the reacted fluid and the other fluid.
  • the reaction module includes a fluid reaction structure and a temperature control component
  • the reaction channel is located on the fluid reaction structure
  • the temperature control component is used to control the temperature of the fluid reaction structure to regulate the temperature in the reaction channel
  • the reaction channel is bent and arranged on the fluid reaction structure.
  • the reaction channel along the extension direction of the reaction channel, the reaction channel includes a plurality of reaction areas, and the temperature control component is further used to control the temperatures of at least two of the reaction areas to be different.
  • the reaction channel is connected to the fluid distribution module through a first connection end, and the reaction channel is connected to the fluid driving module through a second connection end, the first connection end and the second connection end are located at the same end of the fluid reaction structure, or the first connection end and the second connection end are located at opposite ends of the fluid reaction structure.
  • reaction channel, the first connecting end, and the second connecting end are an integrated structure.
  • both the first connection end and the second connection end are provided with a switch component, and the switch component is used to open or close the reaction channel.
  • the switch component includes a solenoid valve or a rotary valve.
  • the fluid reaction structure includes a substrate and a groove formed on the substrate, a hollow reaction pipeline is provided in the groove, and the hollow portion of the reaction pipeline constitutes the reaction channel. Two ends of the reaction pipeline extending out of the groove are respectively communicated with the fluid distribution module and the fluid driving module.
  • the depth of the groove is greater than or equal to the outer diameter of the reaction pipeline.
  • reaction pipeline is detachably disposed in the groove; and/or the reaction pipeline is detachably connected to the fluid distribution module and the fluid driving module.
  • the inner diameter of the reaction pipeline is 0.3 mm to 2 mm.
  • the fluid reaction structure includes a stacked substrate and a cover plate, a groove is formed on the substrate, and a surface of the cover plate close to the substrate is sealed to the substrate so that the groove forms the reaction channel.
  • the fluid reaction structure includes a stacked substrate and a cover plate, and two support walls located between the substrate and the cover plate, and opposite end surfaces of each support wall are respectively connected to the substrate and the cover plate, and the substrate, the two support walls and the cover plate together form the reaction channel.
  • the fluid distribution module includes a liquid collection structure, a distribution pipeline connecting the liquid collection structure and the reaction channel, and a liquid collection drive device, wherein the liquid collection drive device is used to drive the liquid collection structure to align with the fluid storage device and extend into the fluid storage device.
  • the liquid collection structure is a liquid collection needle.
  • the fluid driving module includes a driving pump, and the driving pump is a syringe pump, a plunger pump, a diaphragm pump, a gear pump or a peristaltic pump.
  • the driving pump is a syringe pump, a plunger pump, a diaphragm pump, a gear pump or a peristaltic pump.
  • the sample preparation system further includes a waste liquid collection device and a cleaning liquid storage device, and both the waste liquid collection device and the cleaning liquid storage device are connected to the fluid driving module.
  • the reaction channel is also connected to a sample use system, and the fluid driving module is further used to drive the fluid after reaction in the reaction channel to be transported to the sample use system.
  • a fluid dispensing valve is provided between the fluid dispensing module and the reaction module, and the fluid dispensing valve can also be connected to the sample use system.
  • the sample preparation system further includes a control module, and the control module is used to control the fluid storage module, the fluid distribution module, the fluid driving module, and the reaction module to work in coordination.
  • an embodiment of the present application provides a sample preparation method, comprising:
  • the temperature in the reaction channel is regulated by the reaction module to cause the fluid to undergo a biochemical reaction in the reaction channel, thereby obtaining the sample.
  • the method before the fluid undergoes a biochemical reaction in the reaction channel, the method also includes:
  • the method further comprises:
  • the fluid storage module includes a fluid storage device and a fluid switching device, and the fluid storage device includes a plurality of containers for accommodating the fluid;
  • the method further includes:
  • the fluid dispensing module is aligned with a container storing a fluid in the fluid storage device through coordinated actions of the fluid dispensing module and the fluid switching device;
  • the fluid dispensing module is extended into the container.
  • the method further includes:
  • the fluid dispensing module is aligned with a container of the fluid storage device storing another fluid through coordinated actions of the fluid dispensing module and the fluid switching device;
  • the fluid driving module creates a pressure gradient, so that the reacted fluid in the reaction channel is transferred to a container storing another fluid via the fluid distribution module, so that the another fluid is mixed with the reacted fluid to obtain a mixed fluid;
  • the temperature in the reaction channel is regulated by the reaction module to make the mixed fluid react in the reaction channel until the reaction is completed to obtain the sample.
  • the method further includes:
  • the sample in the reaction channel is transported to a sample using system through the fluid driving module.
  • the method further includes:
  • a pressure gradient is created by the fluid driving module, so that the cleaning liquid in the cleaning liquid storage device enters the reaction flow channel to clean the reaction flow channel, and the waste liquid after cleaning is transported to the waste liquid collection device.
  • an embodiment of the present application provides a biochemical substance analysis system, comprising a sample preparation system and a sample usage system connected to the sample preparation system, wherein the sample preparation system is the sample preparation system as described above, and the fluid driving module is also used to transport the sample located in the reaction channel to the sample usage system.
  • the sample use system includes a sequencing chip, the reaction flow channel and the The fluid driving module is connected to the sequencing chip, and the fluid driving module is used to transport the sample located in the reaction channel to the sequencing chip for sequencing.
  • the sample preparation system provided in the embodiment of the present application performs the processes such as fluid pipetting/mixing and biochemical reactions through the collaborative operation of the fluid storage module, the fluid distribution module, the fluid drive module and the reaction module, thereby realizing one-stop automated sample preparation, with simple process, high sample preparation efficiency, and small fluid loss, saving cost. Moreover, due to the reduction of human intervention, it is possible to avoid the situation where manual operation is prone to errors, which is conducive to improving the accuracy of subsequent biochemical substance analysis.
  • an integrated sample preparation system can be formed by the aforementioned multiple modules, and consumables are few, the equipment cost is low, it can be integrated into the biochemical substance analysis system, and the sample use system can share the device components to achieve seamless connection with the sample use system, which can reduce or even do not need to add additional device components, and further reduce the cost of the overall biochemical substance analysis system.
  • the reaction flow channel is arranged on the fluid reaction structure in a circling manner, and the reaction flow channel with a longer length can be centrally arranged to reduce the occupied space, which is conducive to reducing the volume of the fluid reaction structure, so as to reduce the volume of the sample preparation system and the overall biochemical substance analysis system.
  • FIG. 1 is a schematic diagram of the system architecture of a sample preparation system provided in an embodiment of the present application.
  • FIG. 2 is a schematic diagram of the structure of a sample preparation system provided in an embodiment of the present application.
  • FIG. 3 is a schematic structural diagram of a fluid storage device in a sample preparation system provided in another embodiment of the present application.
  • FIG. 4 is a schematic diagram of the three-dimensional structure of the fluid reaction structure in the sample preparation system shown in FIG. 2 .
  • FIG. 5 is a schematic diagram of a planar structure of a fluid reaction structure in the sample preparation system shown in FIG. 4 .
  • FIG. 6 is a schematic structural diagram of a fluid reaction structure provided in another embodiment of the present application.
  • FIG. 7 is a cross-sectional view along line VII-VII in FIG. 5 .
  • FIG8 is a schematic structural diagram of the fluid reaction structure shown in FIG7 after a cover plate is added.
  • FIG. 9 is a schematic structural diagram of a fluid reaction structure in a sample preparation system in another embodiment of the present application.
  • FIG. 10 is a schematic structural diagram of a fluid reaction structure in a sample preparation system in another embodiment of the present application.
  • FIG. 11 is a system architecture diagram of a biochemical substance analysis system provided in an embodiment of the present application.
  • FIG. 12 is a flow chart of a sample preparation method provided in an embodiment of the present application.
  • Sample preparation system 100 Base plate 321, 321a, 321b Fluid storage module 10 grooves 322, 322a, 322b Fluid storage device 101 Reaction pipeline 323 Fluid switching device 102 Cover plate 324 Container 103 Support wall 325 Container bracket 104 Temperature control assembly 303 Transfer mechanism 105 First connection end 304 Driving mechanism 106 Second connection end 305 Mixing mechanism 107 Switch component 306 Fluid distribution module 20 Fluid drive module 40 Liquid extraction structure 201 Drive pump 401 Distribution line 202 Pump drive assembly 402 Liquid taking drive device 203 Control module 50 Fluid dispensing valve 204 Waste collection device 60 Reaction module 30 Cleaning fluid storage device 70 Reaction channel 301 Biochemical material analysis system 1000 Fluid reaction structure 302,302a,302b Sample using system 200
  • FIG. 1 is a schematic diagram of the system architecture of a sample preparation system 100 in one embodiment of the present application.
  • the sample preparation system 100 is used to complete the preliminary preparation of biological samples, including the steps of transferring, mixing, and biochemical reactions of biological samples and reagents.
  • the biological sample can be a human blood sample, a tissue sample, or a saliva sample, as well as other semi-finished biochemical samples, that is, the sample preparation system 100 can realize the steps of transferring, mixing, and biochemical reactions of biological samples and reagents, complete the biochemical reactions between biological samples and different reagents (for example, but not limited to PCR reactions), and the prepared samples can be used for subsequent biochemical substance analysis such as gene sequencing.
  • the sample preparation system 100 includes a fluid storage module 10, a fluid distribution module 20, a reaction module 30, and a fluid drive module 40.
  • the fluid storage module 10 is used to store fluid, wherein the fluid may include biological samples and biochemical
  • the reaction module 30 has a reaction channel 301, which is connected to the fluid distribution module 20 and the fluid driving module 40 respectively.
  • the fluid driving module 40 is used to create a pressure gradient (for example, to generate negative pressure) so that the fluid in the fluid storage module 10 is transported to the reaction channel 301 via the fluid distribution module 20.
  • the fluid driving module 40 can form a negative pressure in the fluid distribution module 20, thereby sucking the fluid in the fluid storage module 10, and at the same time, a negative pressure is also formed in the reaction channel 301, so that the fluid further enters the reaction channel 301.
  • the reaction module 30 is used to perform a biochemical reaction of the fluid transported to the reaction channel 301.
  • the fluid undergoing the biochemical reaction can include a biological sample and the required reagents.
  • a PCR reaction can be implemented in the reaction channel 301.
  • the fluid driving module 40 is also used to create a pressure gradient (e.g., generate positive pressure) so that the reacted fluid in the reaction channel 301 is transported to the container of the fluid storage module 10 storing another fluid via the fluid distribution module 20.
  • the fluid storage module 10 is also used to generate a driving force so that the other fluid is mixed evenly with the reacted fluid to form a mixed fluid, which is further transported to the reaction channel 301 for reaction.
  • the sample preparation system 100 also includes a control module 50, which is used to control the coordinated operation of the fluid storage module 10, the fluid distribution module 20, the reaction module 30 and the fluid driving module 40.
  • the fluid storage module 10 includes a fluid storage device 101 and a fluid switching device 102.
  • the fluid storage device 101 includes a plurality of containers 103 for accommodating the fluid, usually including a sample container and a plurality of reagent containers.
  • the container 103 may be a test tube or other container of any shape that can be used to load biological samples or reagents. As shown in FIG. 2, the container 103 in this embodiment is a test tube.
  • the opening of the container 103 may but need not be sealed with a puncturable sealing film (not shown).
  • the fluid distribution module 20 can extend into the container 103 to absorb fluid.
  • an identification code (such as a QR code or a bar code, etc., not shown) is affixed to the container 103 containing the biological sample.
  • the identification code records the identification information of the corresponding biological sample, such as the name, age, test items, etc. of the test subject, which is used for tracking and management of the biological sample.
  • the amount of biological samples and reagents in the container 103 can be quantitative. According to the needs of the biochemical reaction, the required amount of biological samples and reagents are placed in the container 103 in advance, respectively, to achieve the purpose of quantitative pipetting. It is understandable that the amount of fluid sucked by the fluid distribution module 20 can also be controlled by controlling the pressure gradient generated by the fluid driving module 40 to achieve the purpose of quantitative pipetting.
  • the fluid storage device 101 can also include a container holder 104, and the container 103 is placed in the container holder 104, and the container holder 104 is placed on the fluid switching device 102, which can achieve the stability of multiple containers 103 and facilitate the aspiration operation of the fluid distribution module 20.
  • the fluid switching device 102 is used to adjust the positions of the aforementioned multiple containers 103 so that one of the multiple containers 103 is aligned with the fluid distribution module 20 to achieve the purpose of transferring the fluid in the container 103.
  • the fluid switching device 102 may include a transfer mechanism 105 and a drive mechanism 106.
  • the multiple containers 103 loaded with biological samples and reagents are arranged in sequence on the container holder 104 in a specific order (e.g., a sample loading order), and the container holder 104 is placed on the transfer mechanism 105.
  • the transfer mechanism 105 is driven by the drive mechanism 106.
  • the above containers 103 are sequentially moved to the bottom of the fluid distribution module 20 under the action of the driving mechanism 106, so that the fluid distribution module 20 can absorb the fluid sequentially.
  • the multiple containers 103 can also be arranged in an array, and the transfer mechanism 105 can achieve multi-directional movement on the horizontal plane under the drive of the driving mechanism 106, so that the multiple containers 103 are moved to the bottom of the fluid distribution module 20 in a predetermined order.
  • the transfer mechanism 105 can be a single-axis slide rail that can realize reciprocating movement in a single direction in the horizontal plane, or it can be a dual-axis slide rail that can realize reciprocating movement in two vertical directions in the horizontal plane, thereby switching the relative positions of different containers 103 in the fluid storage device 101 to facilitate liquid extraction by the fluid distribution module 20.
  • the drive mechanism 106 may be a drive motor.
  • the fluid switching device 102 further includes a mixing mechanism 107, which may be located on the transfer mechanism 105, and the container 103 is located on the mixing mechanism 107.
  • the mixing mechanism 107 may mix the fluid in the container 103 uniformly, for example, by using ultrasound, rotation, oscillation, centrifugation, etc. to achieve the mixing of the fluid in the container 103, that is, the mixing mechanism 107 may be a mechanism that can generate ultrasound, rotation, oscillation, centrifugation, etc.
  • the fluid distribution module 20 includes a liquid collection structure 201, a distribution pipeline 202 and a liquid collection drive device 203.
  • the liquid collection structure 201 and the distribution pipeline 202 are interconnected.
  • the distribution pipeline 202 is also connected to the reaction channel 301, and further connected to the fluid drive module 40.
  • the fluid drive module 40 creates a pressure gradient, which can form a pressure gradient, such as negative pressure or positive pressure, in the reaction channel 301, the distribution pipeline 202 and the liquid collection structure 201, thereby realizing the transfer of fluid.
  • the liquid collection drive device 203 is used to drive the liquid collection structure 201 to align with the fluid storage device 101 and extend into the fluid storage device 101.
  • the liquid collection drive device 203 can drive the liquid collection structure 201 to move up and down, so that the liquid collection structure 201 extends into the container 103 to absorb fluid.
  • the liquid extraction structure 201 may be a reagent needle, which extends into the container 103 and extends below the fluid surface to absorb the fluid.
  • the fluid driving module 40 may include a driving pump 401 and a pump driving assembly 402.
  • the pump driving assembly 402 may enable the driving pump 401 to create a pressure gradient, such as negative pressure or positive pressure, to achieve the purpose of fluid driving. It is understandable that the fluid driving module 40 may also use other structures or devices that can generate a pressure gradient.
  • the reaction channel 301 is bent and arranged on the fluid reaction structure 302.
  • the reaction channel 301 is arranged on the fluid reaction structure 302 in a bent manner, and the reaction channels 301 with longer lengths can be arranged in a centralized manner, thereby reducing the occupied space and facilitating the reduction of the volume of the fluid reaction structure 302.
  • the reaction channel 301 is arranged on the fluid reaction structure 302 in a spiral manner, and as shown in FIG. 6 , the reaction channel 301 is arranged on the fluid reaction structure 302 in an S shape.
  • the reaction channel 301 can also be in other irregular curved shapes.
  • the reaction channel 301 can also be arranged on the fluid reaction structure in a straight line.
  • the reaction channel 301 includes multiple reaction areas, and the temperature control component 303 is also used to control the different temperatures of at least two reaction areas.
  • the reaction channel 301 includes reaction area A, reaction area B and reaction area C, which have different temperatures to achieve different stages of biochemical reactions.
  • different stages of PCR reactions require different temperatures, and the reaction process at different stages can be completed by controlling the fluid droplets to move back and forth between reaction area A, reaction area B and reaction area C.
  • the inner diameter of the reaction channel 301 can be 0.3 mm to 2 mm.
  • the reaction channel 301 can be designed as a slender channel, and the slender reaction channel 301 can be bent and set on the fluid reaction structure 302. Without changing the overall capacity of the reaction channel 301, the contact area between the reactants and the atmosphere can be reduced, the problem of reagent crystallization can be avoided, and manual maintenance of the reaction channel 301 can be avoided, thereby improving the efficiency of sample preparation and the utilization rate of the sample preparation system 100.
  • the reaction module 30 also includes a first connection end 304 connecting the reaction channel 301 to the fluid distribution module 20, and a second connection end 305 connecting the reaction channel 301 to the fluid driving module 40, wherein, as shown in FIG. 5 , the first connection end 304 and the second connection end 305 can be located at the same end of the fluid reaction structure 302, and the reaction channel 301 is spirally wrapped around the fluid reaction structure 302.
  • the first connection end 304 and the second connection end 305 are arranged at the same end of the fluid reaction structure 302, which facilitates the overall circuit layout and connection assembly of the sample preparation system 100, makes the layout of each module more centralized, is conducive to making full use of space, reduces the occupied space of the overall sample preparation system 100, and realizes the compactness and miniaturization of the sample preparation system 100. It is understandable that in other embodiments, as shown in FIG6 , the first connection end 304 and the second connection end 305 can also be located at opposite ends of the fluid reaction structure 302.
  • the first connection end 304 and the second connection end 305 can be arranged at opposite ends of the fluid reaction structure 302 to facilitate pipeline connection and improve space utilization.
  • the first connection end 304 and the second connection end 305 can also be arranged at appropriate positions of the fluid reaction structure 302 according to actual needs.
  • the first connection end 304 and the second connection end 305 are both provided with a switch component 306, and the switch component 306 can be used to open or close the reaction channel 301.
  • a switch component 306 is provided at the first connection end 304 of the reaction channel 301, and the fluid distribution module 20
  • the distribution pipeline 202 of the reaction channel 301 is connected to the switch component 306, and the connection and disconnection between the reaction channel 301 and the distribution pipeline 202 can be achieved by controlling the switch component 306.
  • Another switch component 306 is arranged on the second connection end 305 of the reaction channel 301, and the driving pump 401 in the fluid driving module 40 is connected to the switch component 306, and the connection and disconnection between the reaction channel 301 and the reaction channel 301 can be achieved by controlling the switch component 306.
  • the pump head switching valve of the driving pump 401 can be used as the switch component 306 to achieve the purpose of opening and closing the second connection end 305 of the reaction channel 301.
  • the reaction channel 301 can be sealed and isolated from the atmosphere, thereby avoiding fluid boiling and evaporation losses that affect the quality of the biochemical reaction during high-temperature biochemical reactions (such as high-temperature PCR reactions), especially in high-altitude and low-pressure areas. It can also avoid the need for traditional PCR reactions to add a PCR hot cover and the need to manually maintain reagent crystallization on the PCR hot cover.
  • the switch component 306 may be a device component that can realize the opening and closing of the reaction channel 301, for example, it may be a device component such as a solenoid valve, a rotary valve, a metering pump, a plug, or a clamping component such as a stop clamp, etc.
  • the switch components 306 provided at both ends of the reaction channel 301 are stop valves or switching valves.
  • the fluid reaction structure 302 includes a substrate 321 and a groove 322 formed on the substrate 321, a hollow reaction pipeline 323 is arranged in the groove 322, the hollow part of the reaction pipeline 323 constitutes the reaction channel 301, and the two ends of the reaction pipeline 323 extending out of the groove 322 respectively constitute the first connection end 304 and the second connection end 305, thereby realizing communication with the fluid distribution module 20 and the fluid driving module 40.
  • the reaction pipeline 323 can be a slender and flexible hose, and can be made of Teflon materials such as FEP, PFA, PTFE or other materials that are not easily corroded and have a certain flexibility.
  • the overall length of the groove 322 on the substrate 321 can be designed according to the actual required length of the reaction channel 301, and the bending mode of the groove 322 can be reasonably set according to the area of the substrate 321, so that the slender reaction channel 301 is set on the substrate 321, which is convenient for the bending setting of the reaction channel 301 on the substrate 321.
  • the groove 322 can be formed by etching, laser, mechanical or injection molding, and the curved shape of the groove 322 can be flexibly set, which is conducive to making full use of the space of the substrate 321, thereby reducing the volume of the reaction module 30.
  • reaction pipeline 323 when the flow rate is small, the length of the required reaction pipeline 323 is short, and the reaction pipeline 323 can also be designed in a straight line, or according to assembly requirements, in order to reasonably use the space, the reaction pipeline 323 can also be set on the fluid reaction structure 302 in a straight line.
  • the fluid reaction structure 302 may have a heat conduction function, and the temperature control component 303 may directly control the temperature of the fluid reaction structure 302, thereby achieving temperature control of the reaction pipeline 323 embedded in the groove 322, so as to accurately control the temperature in the reaction channel 301.
  • the temperature control component 303 may be a thermoelectric cooler (TEC), which can achieve heating and cooling of the substrate 321.
  • TEC thermoelectric cooler
  • the depth of the groove 322 (such as L2 in FIG. 7 ) is greater than or equal to the outer diameter of the reaction tube 323 (such as L1 in FIG. 7 ).
  • the reaction tube 323 can be completely embedded in the groove 322, so that the temperature in the reaction tube 323 is more uniform. uniformity to improve the accuracy of temperature control in the reaction channel 301.
  • the reaction pipeline 323 is detachably disposed in the groove 322.
  • the hose of the reaction pipeline 323 can be directly embedded in the groove 322, which is convenient and fast, and easy to maintain and replace.
  • the reaction channel 301, the first connection end 304 and the second connection end 305 are an integrated structure, and the passages of the above three parts are in the form of fluid pipelines to form an integrated design, so that the reaction area, the pipetting needle and the pump valve and other connecting pipelines in the reaction module 30, the fluid distribution module 20 and the fluid drive module 40 are all on the same fluid channel.
  • the PCR area and the pump valve fluid pipeline share a pipeline.
  • the above design greatly reduces the sample loss, the number of consumables is small, and the process operation is simple.
  • the reaction pipeline 323 can also be detachably connected to the fluid distribution module 20 and the fluid drive module 40, which is convenient for separate maintenance or replacement of each part.
  • the reaction line 323 may be a non-consumable material, and the reaction line 323 may be reused by cleaning the reaction channel 301, effectively reducing the cost of the sample preparation system 100. It is understandable that the reaction line 323 may also be a consumable material, and the reaction line 323 is replaced after each sample preparation, without the need for a cleaning process, and the reaction line 323 is easy to replace.
  • the sample processing volume of the reaction channel 301 is flexible, and the sample processing volume can be controlled by designing the inner diameter and length of the reaction channel 301.
  • the reaction channel 301 of the sample preparation system 100 of this embodiment can achieve a sample processing volume of 3 ⁇ L to 2500 ⁇ L, or a wider range of sample processing volumes.
  • the fluid reaction structure 302 further includes a cover plate 324 covering the substrate 321 , and the cover plate 324 covers the groove 322 .
  • the addition of the cover plate 324 can make the reaction pipeline 323 in the groove 322 be heated more evenly, and in particular can make the heating conditions of the reaction pipeline 323 at the opening of the groove 322 and the reaction pipeline 323 at the bottom of the groove 322 consistent.
  • the reaction channel 301 can also be directly formed on the fluid reaction structure.
  • the fluid reaction structure 302a can also include a stacked substrate 321a and a cover plate 324, a groove 322a is formed on the substrate 321a, and the surface of the cover plate 324 close to the substrate 321a is sealed and connected to the substrate 321a so that the groove 322a forms a sealed reaction channel 301. That is, in this embodiment, the groove 322a is directly formed on the substrate 321a to form the reaction channel 301, so that the reaction module 30 structure is simpler, and the temperature control of the reaction channel 301 formed by the groove 322a is easier, and the temperature is more uniform.
  • the groove 322a can be obtained by injection molding or etching.
  • the fluid reaction structure 302b includes a stacked substrate 321b and a cover plate 324, and two support walls 325 located between the substrate 321b and the cover plate 324.
  • the two opposite end surfaces of each support wall 325 are respectively connected to the substrate 321b and the cover plate 324.
  • the substrate 321b, the two support walls 325 and the cover plate 324 together enclose the reaction channel 301.
  • the volume of the reagent reaction channel 301 can be reduced by designing the thickness and spacing of the two support walls 325.
  • the two support walls 325 can be made of a flexible material.
  • the reaction channel 301 may be designed according to the desired curved shape.
  • the pipeline when the fluid reaction structure is connected with the fluid distribution module 20 and the fluid driving module 40, the pipeline can be inserted into the port of the reaction channel 301 corresponding to the fluid reaction structure 302a or 302b to achieve communication.
  • the reaction channel 301 is directly formed on the fluid reaction structure 302a or 302b, the overall fluid reaction structure 302a or 302b as a whole can be detachably connected with the distribution pipeline 202 in the fluid distribution module 20 and the distribution pipeline 202 in the fluid driving module 40, which is convenient for maintenance and replacement of the fluid reaction structure 302a or 302b.
  • the sample preparation system 100 also includes a waste liquid collection device 60 and a cleaning liquid storage device 70, and both the waste liquid collection device 60 and the cleaning liquid storage device 70 are connected to the fluid drive module 40. Specifically, the waste liquid collection device 60 and the cleaning liquid storage device 70 are both connected to the drive pump 401 in the fluid drive module 40.
  • the drive pump 401 is a syringe pump
  • the waste liquid collection device 60 and the cleaning liquid storage device 70 are connected to the pump head of the syringe pump through a pipeline.
  • pure water can be placed in the cleaning liquid storage device 70 to clean the entire flow channel.
  • the processes such as fluid pipetting, mixing and biochemical reaction can be performed, so that one-stop automated sample preparation (such as library processing before sequencing) can be realized, the process is simple, the sample preparation efficiency is high, and the fluid loss is small, which saves costs and realizes low-cost small modular design; moreover, the aforementioned multiple modules can constitute an integrated sample preparation system, so that the reaction flow channel 301 is connected with other fluid pipelines to form an integrated fluid channel, the sample loss is reduced, the consumables are small, the equipment cost is low, and the operation process is simple.
  • the sample preparation system 100 can be integrated into a sample use system (such as a sequencer), and some components in the sample use system can be shared, and the flux matching is better.
  • the sample preparation system 100 can avoid the situation where manual operation is prone to errors due to reduced human intervention, which is beneficial to improving the accuracy of subsequent biochemical substance analysis.
  • the fluid is confined in the reaction channel 301 to achieve front-to-back sealing, thereby avoiding liquid boiling and evaporation loss during high-temperature biochemical reactions, especially in high-altitude and low-pressure areas; after the reaction channel 301 is sealed, the reactants are isolated from the atmosphere, and the gas-liquid interface is greatly reduced or even eliminated, thereby solving the problem of PCR quality degradation caused by PCR high-temperature evaporation and improving PCR quality; the fluid is isolated from the atmosphere, and the problem of adding a "PCR hot cover" commonly seen in traditional automated sample preparation systems and manual maintenance of the "PCR hot cover” can be avoided.
  • the reaction channel 301 is embedded in the fluid reaction structure 302 that can achieve temperature change, which facilitates the temperature control of the reaction channel 301.
  • the reaction channel 301 can be designed as “non-consumable”, reducing the cost of sample preparation.
  • the reaction channel 301 can also be designed as a consumable, which is easy to replace and does not require cleaning, further simplifying the operation and structure of the sample preparation system 100.
  • the sample preparation system 100 has a flexible sample processing volume, and can process samples in a range of 3 ⁇ L to 2500 ⁇ L, or a wider range.
  • the embodiment of the present application further provides a biochemical substance analysis system 1000.
  • the biochemical substance analysis system 1000 includes the sample preparation system 100 as described above and a sample use system 200 connected to the sample preparation system 100.
  • the fluid driving module 40 in the sample preparation system 100 is also used to transfer the product sample obtained after the reaction in the reaction channel 301 to the sample use system 200, that is, the fluid driving module 40 can create a pressure gradient to drive the product sample obtained after the reaction in the reaction channel 301 to be transferred to the sample use system 200.
  • the sample preparation system 100 can be directly integrated with the sample use system 200 to directly transfer the product sample prepared in the reaction channel 301 to the sample use system 200 for subsequent biochemical substance analysis process.
  • the sample preparation system 100 can realize PCR amplification of biological samples
  • the sample use system 200 can be a sequencer, including a sequencing chip for realizing gene sequencing.
  • the reaction channel 301 of the sample preparation system 100 is connected to the sequencing chip of the sample use system 200, and the fluid driving module 40 transfers the product sample obtained by PCR reaction in the reaction channel 301 to the sequencing chip for sequencing.
  • a fluid distribution valve 204 is further provided between the fluid distribution module 20 and the reaction module 30.
  • the fluid distribution valve 204 can also be connected to the sample use system 200.
  • the fluid distribution valve 204 can be a solenoid valve, having a normally open end, a common end and a normally closed end.
  • the fluid distribution module 20 is connected to the normally open end
  • the reaction module 30 is connected to the common end
  • the sample use system 200 is connected to the normally closed end.
  • the normally closed end is opened and the normally open end is closed.
  • the distribution pipeline 202 is connected to the normally open end
  • the first connection end 304 of the reaction flow channel 301 is connected to the common end
  • the sequencing chip of the sample use system 200 is connected to the normally closed end.
  • the sample preparation system 100 of the embodiment of the present application can be integrated with the conventional sample use system 200, can share components (such as pumps, valves, pipelines, drivers, control software, UI interface, etc.) with the sample use system 200, and can be integrated into the sample use system 200 without adding or adding less other components. It can achieve seamless connection with the sample use system 200, can reduce or even eliminate the need to add additional components, further reduce the cost of the overall biochemical substance analysis system 1000, significantly shorten working time, and reduce human errors.
  • components such as pumps, valves, pipelines, drivers, control software, UI interface, etc.
  • the embodiment of the present application further provides a method for preparing a product sample using the aforementioned sample preparation system 100 , which specifically includes the following steps:
  • step S01 a pressure gradient is created by the fluid driving module 40 , so that the fluid in the fluid storage module 10 is transported to the reaction channel 301 in the reaction module 30 via the fluid distribution module 20 .
  • Step S02 regulating the temperature in the reaction channel 301 through the reaction module 30 to cause the fluid to undergo a biochemical reaction in the reaction channel 301 , thereby obtaining the sample, ie, the product sample.
  • the method for preparing the product sample includes the following steps:
  • Step S1 through the coordinated action of the fluid distribution module 20 and the fluid switching device 102, the fluid distribution The module 20 is aligned with a container 103 storing a first fluid in the fluid storage device 101 .
  • Step S2 extending the fluid distribution module 20 into the container 103 storing the first fluid.
  • Step S3 creating a pressure gradient through the fluid driving module 40 , so that the first fluid in the container 103 storing the first fluid is transported to the reaction channel 301 in the reaction module 30 through the fluid distribution module 20 .
  • Step S4 closing the switch components 306 at both ends of the reaction channel 301 to seal the reaction channel 301 .
  • Step S5 regulating the temperature in the reaction channel 301 through the reaction module 30 , so that the first fluid undergoes a biochemical reaction in the reaction channel 301 .
  • Step S6 opening the switch components 306 at both ends of the reaction channel 301.
  • Step S7 through the coordinated action of the fluid dispensing module 20 and the fluid switching device 102 , the fluid dispensing module 20 is aligned with the container 103 storing the second fluid of the fluid storage device 101 .
  • Step S8 extending the fluid distribution module 20 into the container 103 storing the other fluid.
  • Step S9 creating a pressure gradient through the fluid driving module 40, so that the first fluid after reaction in the reaction channel 301 is transferred to the container 103 storing the second fluid through the fluid distribution module 20, so that the second fluid is mixed with the first fluid after reaction to obtain a mixed fluid.
  • step S10 a pressure gradient is created by the fluid driving module 40 so that the fluid distributing module 20 absorbs the mixed fluid and transfers the mixed fluid to the reaction channel 301 in the reaction module 30 .
  • step S11 the temperature in the reaction channel 301 is regulated by the reaction module 30 to allow the mixed fluid to react in the reaction channel 301 until the reaction is completed to obtain a product sample.
  • the method further comprises:
  • the sample in the reaction channel 301 is transported to the sample using system through the fluid driving module 40 .
  • the method further includes:
  • the pressure gradient is created by the fluid driving module 40 , so that the cleaning liquid in the cleaning liquid storage device 70 enters the reaction channel 301 to clean the reaction channel 301 , and the waste liquid after cleaning is transferred to the waste liquid collection device 60 .
  • the control module 50 includes a control program for controlling the coordinated operation of the fluid storage module 10, the fluid distribution module 20, the fluid driving module 40 and the fluid reaction module 30.
  • the control program When the control program is running, it is used to execute the following method:
  • Step 1 The transfer mechanism 105 in the fluid switching device 102 coordinates with the liquid extraction drive device 203 to align the needle tip of the liquid extraction structure 201 (such as a reagent needle) in the vertical direction with the container (sample container) storing the first fluid, ready to absorb the first fluid, where the first fluid can be a biological sample, such as a DNA sample.
  • the liquid extraction structure 201 such as a reagent needle
  • Step 2 The reagent needle is lowered and inserted below the liquid level of the sample tube, driving the pump 401 (such as a syringe pump) to operate and guide the first fluid into the reaction channel 301 .
  • the pump 401 such as a syringe pump
  • Step 3 The temperature control component 303 controls the temperature of the fluid reaction structure 302, so that the first fluid can achieve different temperature reactions (such as PCR reactions) in the reaction channel 301. Close the switch component 306 so that the fluid in the reaction channel 301 is isolated from the atmosphere. After the temperature control is completed, open the switch component 306.
  • Step 4 The transfer mechanism 105 and the liquid extraction drive device 203 work in coordination to align the needle tip of the reagent needle with the container 103 containing the second fluid in the vertical direction.
  • Step 5 Control the syringe pump to control the vertical movement of the reagent needle, guide the first fluid after the reaction in the reaction channel 301 to the reagent needle, and discharge it into the container 103 containing the second fluid.
  • Step 6 The mixing mechanism 107 mixes the first fluid and the second fluid after the reaction in the container 103 to obtain a mixed fluid, for example, by using methods such as ultrasound, rotation, vibration, centrifugation, etc.
  • Step 7 The syringe pump is activated to control the movement of the reagent needle in the vertical direction and guide the mixed fluid into the reaction channel 301 for the second step reaction.
  • Step 8 Repeat steps 3 to 7 to achieve the steps of mixing multiple fluids, quantitative pipetting, biochemical reaction in a closed micro-chamber, etc., and finally obtain a product sample for subsequent biochemical substance analysis (such as gene sequencing).

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Abstract

L'invention concerne un module de réaction, un système de préparation d'échantillon, un procédé de préparation d'échantillon et un système d'analyse de substance biochimique. Le système de préparation d'échantillon comprend un module de stockage de fluide, un module de distribution de fluide, un module d'entraînement de fluide et un module de réaction, le module de stockage de fluide étant utilisé pour stocker un fluide; le module de réaction a un canal d'écoulement de réaction, et le canal d'écoulement de réaction est en communication avec à la fois le module de distribution de fluide et le module d'entraînement de fluide; le module d'entraînement de fluide est utilisé pour générer un gradient de pression, de telle sorte que le fluide dans le module de stockage de fluide est transféré au canal d'écoulement de réaction par l'intermédiaire du module de distribution de fluide; et le module de réaction est utilisé pour exécuter une réaction biochimique du fluide. Dans la présente invention, grâce à la collaboration entre le module de stockage de fluide, le module de distribution de fluide, le module d'entraînement de fluide et le module de réaction, le transfert/mélange et une réaction biochimique du fluide sont mis en oeuvre, ce qui permet de réaliser une préparation automatique d'échantillon en une seule étape. La présente invention fait intervenir un processus simple, présente des niveaux élevés d'efficacité et une perte de fluide relativement faible, réduit les coûts et présente une réaction biochimique de haute qualité.
PCT/CN2023/117644 2023-09-08 2023-09-08 Module de réaction, système de préparation d'échantillon, procédé de préparation d'échantillon et système d'analyse de substance biochimique Pending WO2025050373A1 (fr)

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PCT/CN2023/117644 WO2025050373A1 (fr) 2023-09-08 2023-09-08 Module de réaction, système de préparation d'échantillon, procédé de préparation d'échantillon et système d'analyse de substance biochimique

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011156854A1 (fr) * 2010-06-17 2011-12-22 Geneasys Pty Ltd Dispositif microfluidique doté d'un capteur de conductivité
CN107988044A (zh) * 2017-12-29 2018-05-04 东南大学 一种大反应体积流道式pcr扩增装置
CN110560184A (zh) * 2018-06-06 2019-12-13 厦门大学 微流控芯片、微流控反应系统及驱动方法
KR20210015292A (ko) * 2019-08-01 2021-02-10 한국과학기술원 모듈형 미세 유체 장치 및 이의 제조 방법
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CN113122613A (zh) * 2021-04-09 2021-07-16 四川微康朴澜医疗科技有限责任公司 一种全封闭式荧光定量pcr微流控检测芯片

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