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WO2025011622A1 - Support - Google Patents

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Publication number
WO2025011622A1
WO2025011622A1 PCT/CN2024/105017 CN2024105017W WO2025011622A1 WO 2025011622 A1 WO2025011622 A1 WO 2025011622A1 CN 2024105017 W CN2024105017 W CN 2024105017W WO 2025011622 A1 WO2025011622 A1 WO 2025011622A1
Authority
WO
WIPO (PCT)
Prior art keywords
wall
cavity
sealing
carrier according
carrier
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/CN2024/105017
Other languages
English (en)
Chinese (zh)
Inventor
韦嘉
徐强
赵蒙
徐涛
冼志科
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.)
Guangzhou National Laboratory
Original Assignee
Guangzhou National Laboratory
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 Guangzhou National Laboratory filed Critical Guangzhou National Laboratory
Publication of WO2025011622A1 publication Critical patent/WO2025011622A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • 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
    • C12M1/02—Apparatus for enzymology or microbiology with agitation means; with heat exchange means
    • 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
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • 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 field of in vitro diagnostic technology, and in particular to a carrier.
  • PCR polymerase chain reaction
  • the reaction sample Before PCR amplification, the reaction sample needs to be placed in a carrier, where the reaction sample consists of samples such as collected throat swabs or nasal swabs and reaction samples used for PCR amplification.
  • the reaction sample Before PCR amplification, the reaction sample needs to be heated by a heater and cooled by a cooling mechanism, so that the reaction sample cycles through high temperature denaturation, low temperature annealing and suitable temperature extension stages.
  • Some carriers in the prior art are tube structures such as ep tube structures.
  • the diameter of the area where the carrier holds the reaction sample is large, the heat-averaging speed is slow, the reaction sample temperature rise and fall speed is slow, and the reaction tube is difficult to fit tightly with the heating structure, which affects the temperature rise and fall speed.
  • the diameter of the area where the tube carrier holds the reaction sample is large, the heat-averaging speed of the reaction sample is slow, and the reaction sample temperature rise and fall speed is slow.
  • the wall thickness of the carrier with a tube structure is large, and the heat transfer is slow, which further leads to the slow temperature rise and fall speed of the reaction sample.
  • the reaction sample is injected into the sample holding cavity through the opening of the PCR tube, and the gas in the sample holding cavity is also discharged from the opening.
  • the injection port in the carrier is small, the gas is difficult to discharge, resulting in a large amount of gas remaining in the carrier, thereby affecting the amplification efficiency.
  • the gas remaining in the carrier and the gas precipitated in the reaction sample during the amplification process are often mixed in the reaction sample, thereby affecting the detection results and affecting the amplification efficiency.
  • the purpose of this application is to propose a carrier to solve at least one of the above technical problems.
  • the present application provides a carrier, including a main body and a sealing member, wherein the main body is provided with a sample holding cavity and a sealing cavity, the sample holding cavity is connected to the sealing cavity, at least part of the cavity wall of the sample holding cavity is a deformable wall, at least part of the sealing member can be sealably inserted into the sealing cavity to at least compress the gas in the sealing cavity, and make the deformable wall protrude outward so that the deformable wall is in close contact with a temperature regulating device.
  • the deformable wall is a deformable membrane.
  • the sealing member can be inserted into the sealing cavity in a sealing manner to compress the gas in the sealing cavity and the sample holding cavity.
  • the main body and/or the sample holding cavity is a flat structure.
  • the body includes a first wall and a second wall arranged opposite to each other, and a and a side wall between the first wall and the second wall, the first wall and/or the second wall comprising the deformable wall.
  • At least positions of the first wall and the second wall opposite to the sample holding cavity are both the deformable walls.
  • the side wall is made of transparent material.
  • At least the position of the first wall opposite to the sample holding chamber is the deformable wall.
  • the second wall and/or the side wall are made of a transparent material.
  • the second wall and the side wall are integrally formed.
  • the thickness of the second wall is not greater than 0.5 mm.
  • the sealing member includes a first plug, and the first plug is sealingly disposed in the sealing cavity.
  • the sealing member comprises a second plug column and a sealing portion connected to each other, wherein the sealing portion is arranged around the outer circumference of the second plug column and is spaced apart from the outer circumference of the second plug column, and the outer circumference of the sealing portion can be in sealing contact with the cavity wall of the sealing cavity.
  • a sealing convex ring is formed on the outer periphery of the sealing portion, and the sealing convex ring is in sealing contact with the cavity wall of the sealing cavity.
  • the sealing member further comprises a positioning cap, and ends of the second plug and the sealing portion are both connected to the positioning cap, and the positioning cap can abut against an outer surface of the body.
  • the seal is integrally formed with the body.
  • the main body further includes an annular wall, the annular wall is connected to the side wall, and the space inside the annular wall forms the sealed cavity.
  • the sealing member is interference fit with the sealing cavity.
  • the flat structure means that the dimension of the sample holding cavity or the body in a direction perpendicular to its thickness direction is larger than its dimension in the thickness direction.
  • the ratio of the dimension of the sample holding cavity or the body in a direction perpendicular to its thickness direction to its thickness direction is greater than 5:1.
  • the ratio of the dimensions is 50:1 to 100:1.
  • a liquid inlet channel is further provided on the main body, and the sample containing cavity and the sealing cavity are communicated with each other through the liquid inlet channel.
  • the upper end of the liquid inlet channel is connected to the sealed cavity, and the lower end of the liquid inlet channel is connected to the lower end of the sample holding cavity.
  • an exhaust channel is further provided on the main body, one end of the exhaust channel is communicated with the sealed cavity, and the other end of the exhaust channel is communicated with the sample holding cavity.
  • the lower end of the exhaust channel is connected to the upper end of the sample holding cavity, and the upper end is connected to the sealed cavity.
  • a liquid injection hole is provided between the liquid inlet channel and the sealing cavity.
  • the inner wall of the injection hole is a truncated cone structure, the large diameter end of the injection hole is connected to the sealing cavity, and the small diameter end is connected to the liquid inlet channel.
  • the deformable wall is an aluminum film, a polypropylene film or a polycarbonate film; or the deformable wall includes an aluminum film and an isolation film, and the isolation film is connected to the inner side of the aluminum film.
  • the thickness of the deformable wall is not greater than 100 microns.
  • the thickness of the isolation film is not greater than 50 microns.
  • the outer surface of the second wall is connected with reinforcing ribs.
  • the carrier further includes a reinforcement portion, and the reinforcement portion is used to reinforce the body.
  • the reinforcement portion includes a first reinforcement portion, and the first reinforcement portion is a strip structure.
  • the main body is a rectangular parallelepiped structure
  • the first reinforcement part is provided on one side or both sides along the width direction of the main body, and the thickness of the first reinforcement part is greater than the thickness of the main body.
  • the reinforcement part further includes a second reinforcement part, and the second reinforcement part is provided on one side or both sides in the length direction of the main body, and the thickness of the second reinforcement part is greater than the thickness of the main body.
  • the carrier further includes a handle portion, and the handle portion is connected to one end of the body.
  • the carrier further includes a positioning portion for positioning the carrier.
  • the positioning portion is a positioning notch opened in the main body or the reinforcement portion.
  • the technical solution provided by the present application is that at least part of the sealing member is sealed and inserted into the sealed cavity.
  • the sealing member occupies the space in the body, the gas in the body is compressed, so that the pressure in the body is greater than the external pressure, and the deformable wall protrudes outward.
  • the temperature regulating device is arranged outside the deformable wall, the deformable wall is in close contact with the temperature regulating device due to the protrusion of the deformable wall, and the heat transfer speed between the deformable wall and the temperature regulating device is fast, and the reaction sample is heated and cooled quickly.
  • the temperature regulating device applies enough pressure to the carrier to press the outer surface of the deformable wall into a plane, the carrier is completely compressed at this time, and the temperature regulating device is in full contact with the carrier, ensuring rapid heat transfer between the temperature regulating device and the carrier, and improving the heating and cooling speed of the reaction sample and the amplification efficiency.
  • the volume of the sample holding cavity is reduced relative to the deformable wall protruding outward, therefore, the gas in the body is further compressed, the air pressure of the sample holding cavity is further increased, and the reaction sample will be in better contact with the deformable wall, which is further conducive to temperature transfer.
  • the internal pressure of the sample holding chamber is increased, so that the deformable wall and the temperature regulating device form good thermal contact, thereby improving the heat conduction efficiency between the deformable wall and the temperature regulating device, and improving the amplification efficiency of the reaction sample.
  • the carrier provided in the present application does not require additional gas to be filled into the carrier or more reaction samples to be introduced into the carrier. Instead, the internal pressure of the sample holding cavity can be adjusted by sealing the seal in the sealing cavity so that the carrier fits with the temperature adjustment device.
  • the reaction sample in the body or sample holding cavity of the flat structure is thin, the center of the reaction sample is very close to the surface of the liquid, the temperature of the reaction sample can reach a uniform temperature in a very short time, and the flat structure can make the contact area between the reaction sample and the temperature regulating device large, the heat transfer efficiency is high, so that the reaction sample temperature rise and fall speed and detection efficiency are greatly improved.
  • the inner diameter of the tube carrier is larger than that of the sample holding cavity of the flat structure, the center of the reaction sample is very close to the surface of the liquid, the temperature of the reaction sample takes a long time to reach a uniform temperature, the reaction sample temperature rise and fall speed is low, and the detection efficiency is low.
  • the main body is provided with an exhaust channel, through which the air in the sample holding chamber can be discharged, thereby avoiding residual air in the sample holding chamber, thereby ensuring rapid equalization of the temperature and temperature rise and fall of the reaction sample.
  • the carrier or sample holding cavity has a flat structure.
  • the carrier or sample holding cavity is arranged vertically, so that the gas released by the reaction sample in the sample holding cavity during the heating process in the amplification stage is more likely to float to the upper end of the main body instead of being dispersed everywhere in the sample holding cavity.
  • the carrier is placed vertically, and the liquid undergoes thermal convection, which is conducive to further mixing the liquid and improving the temperature uniformity of the reaction sample.
  • FIG. 1a is a schematic diagram of the structure of a body provided in an embodiment of the present application.
  • FIG1b is a schematic structural diagram of a sealing member provided in an embodiment of the present application when the sealing member is not covered on the body and the body is unloaded;
  • FIG1c is a schematic structural diagram of a sealing member provided in an embodiment of the present application when the sealing member is not covered on the body and the body is fully loaded;
  • FIG1d is a schematic diagram of a structure in which a sealing member covers a main body and the main body is fully loaded, provided in an embodiment of the present application;
  • FIG. 1e is a schematic structural diagram of a carrier provided in an embodiment of the present application when it is clamped between two sets of temperature adjustment devices;
  • FIG2a is a schematic diagram of the structure of another body provided in an embodiment of the present application.
  • FIG2b is a schematic structural diagram of a sealing member provided in an embodiment of the present application when the sealing member is not covered on another body and the body is unloaded;
  • FIG2c is a schematic structural diagram of a sealing member provided in an embodiment of the present application when the sealing member is not covered on another body and the body is fully loaded;
  • FIG2d is a schematic diagram of a structure in which a sealing member provided in an embodiment of the present application covers another body and the body is fully loaded;
  • FIG2e is a schematic diagram of a structure in which a carrier and a temperature adjustment device are in contact and pressed together according to an embodiment of the present application;
  • FIG3a is an exploded view of a temperature regulating device provided in an embodiment of the present application.
  • FIG3 b is a three-dimensional view of a sealing member provided by an embodiment of the present application when the sealing member is covered on the body and the first wall of the carrier is removed;
  • FIG4 is a perspective view of the sealing member provided in the embodiment of the present application when the sealing member is not covered on the body and the first wall of the carrier is removed;
  • FIG5 is a cross-sectional view of a carrier when the sealing member provided in an embodiment of the present application is not covered on the body;
  • FIG6 is a partial enlarged view of point A in FIG5 ;
  • FIG7 is a cross-sectional view of a carrier when a sealing member provided in an embodiment of the present application covers a body;
  • FIG8 is a partial enlarged view of point B in FIG7;
  • FIG9 is a three-dimensional view of the carrier from another perspective when the sealing member provided by an embodiment of the present application covers the body;
  • FIG10 is a schematic diagram of the structure of a temperature adjustment device provided in an embodiment of the present application.
  • FIG. 11 is a schematic structural diagram of a carrier insertion temperature adjustment device provided in an embodiment of the present application.
  • a first feature being “above” or “below” a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them.
  • a first feature being “above”, “above” and “above” a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature.
  • a first feature being “below”, “below” and “below” a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.
  • connection should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements.
  • connection can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements.
  • This embodiment provides a carrier, which can be used to hold reaction samples that need to be amplified, but is not limited thereto and can also be used in other occasions.
  • the carrier provided in this embodiment includes a main body 1 and a sealing member 2.
  • the main body 1 is provided with a sample holding cavity 14 and a sealing cavity 15.
  • the sample holding cavity 14 is connected to the sealing cavity 15.
  • At least part of the sealing member 2 can be sealably inserted into the sealing cavity 15.
  • At least part of the cavity wall of the sample holding cavity 14 is a deformable wall.
  • the body 1 includes a first wall 11 and a second wall 12 arranged opposite to each other, and a side wall 13 arranged between the first wall 11 and the second wall 12.
  • the first wall 11 and the second wall 12 include deformable walls, or as shown in Figure 2a, the first wall 11 includes a deformable wall.
  • the gas in the sealing chamber 15 is compressed, or when the sample holding chamber 14 carries a reaction sample, but the reaction sample does not fill the sample holding chamber 14, the gas in the sealing chamber 15 and the sample holding chamber 14 is compressed, so that the pressure in the body 1 is greater than the external pressure (usually atmospheric pressure), thereby causing the deformable wall to protrude outward.
  • the external pressure usually atmospheric pressure
  • the temperature regulating device 100 may include a heater 101 and/or a cooler 102, such as the heater 101 is in direct contact with the cooler 102, the carrier is in contact with the heater 101, the heater 101 heats the reaction sample, and the cooler 102 cools the heater 101, thereby cooling the reaction sample, and then realizing PCR amplification.
  • the cooler 102 can also be connected to the plug block 103, and the plug block 103 is provided with a slot, and the carrier can be inserted into the slot, so as to limit the carrier.
  • the deformable wall will slightly protrude. If the temperature regulating device 100 applies enough pressure to the carrier to press the outer surface of the deformable wall into a plane, the carrier is completely compressed at this time, and the temperature regulating device 100 is in full contact with the carrier, ensuring rapid heat transfer between the temperature regulating device 100 and the carrier, and improving the heating and cooling speed of the reaction sample and the amplification efficiency.
  • the volume of the sample holding chamber 14 is reduced relative to when the deformable wall protrudes, so the gas in the body 1 is further compressed, and the gas pressure of the sample holding chamber 14 is further increased than that in Fig. 1d and Fig. 2d, and the temperature regulating device 100 will be in better contact with the deformable wall, which is further conducive to temperature transfer.
  • the volume of the sample holding chamber 14 is designed to be V1, the volume of the sealed chamber 15 is V2, and the volume of the seal 2 inserted into the sealed chamber 15 is V3; as shown in Figures 1c and 2c, after adding a fixed volume of reaction sample V1 into the sample holding chamber 14, the reaction sample fills the sample holding chamber 14; after the seal 2 is sealed and covered on the sealed chamber 15 ( Figures 1d and 2d), the seal 2 compresses the gas with a volume of V3, so that the deformable wall of the sample holding chamber 14 bulges, and the reaction sample level must drop at this time; as shown in Figures 1e and 2e, after applying sufficient pressure to completely press the temperature regulating device 100 and the carrier, the reaction sample will completely fill the sample holding chamber 14 again.
  • the gas is further compressed, and the air pressure of the sample holding chamber 14 is further increased than that of Figures 1d and 2d.
  • the temperature regulating device 100 will be in better contact with the deformable wall, which is conducive to temperature transfer.
  • the carrier provided in this application is not There is no need to inject additional gas into it and there is no need to introduce more reaction samples into the carrier. Instead, the internal pressure of the sample holding chamber 14 can be adjusted by sealing the seal in the sealing cavity so that the carrier fits with the temperature adjustment device 100.
  • the seal 2 can be completely inserted into the sealed cavity 15, or partially inserted into the sealed cavity 15.
  • the sealed cavity 15 can be completely occupied by the seal 2, or not completely occupied (as shown in Figures 7 and 8).
  • the volume of the sealed cavity 15 and the seal 2 can be reasonably designed.
  • the design principle is: after the seal 2 is sealed and inserted into the sealed cavity 15, when the temperature regulating device 100 flattens the deformable wall (at this time, if the filling amount of the reaction sample is consistent with the holding amount of the sample holding cavity 14, the reaction sample can completely fill the sample holding cavity 14), the sample holding cavity 14 forms a positive pressure with the outside (usually atmospheric pressure), for example: the pressure difference between the sample holding cavity 14 and the outside is 0.5 bar, 1 bar, 1.5 bar, etc., but the pressure difference is not limited to this.
  • the internal pressure of the sample holding chamber 14 is increased by reasonably setting the volume of the sample holding chamber 14, the volume of the sealing chamber 15, and the volume of the sealing member 2 inserted into the sealing chamber 15, so that the deformable wall and the temperature regulating device 100 form a good thermal contact, thereby improving the heat conduction efficiency between the deformable wall and the temperature regulating device 100 and improving the amplification efficiency of the reaction sample.
  • the deformable wall in order to further improve the heat conduction rate between the deformable wall and the temperature regulating device 100 while ensuring the strength of the deformable wall, is a deformable membrane.
  • the thickness of the deformable wall is not greater than 100 microns.
  • the deformable wall can be an aluminum film, a polypropylene film or a polycarbonate film; or the deformable wall includes an aluminum film and an isolation film, the isolation film is connected to the inner side of the aluminum film, and the isolation film is directly in contact with the reaction sample to separate the aluminum film and the reaction sample to prevent the aluminum film from affecting the reaction sample.
  • the thickness of the deformable wall is not more than 100 microns, so it can have sufficient strength.
  • the isolation film can be a polypropylene film or a polycarbonate film. The thickness of the isolation film is not more than 50 microns, such as not more than 30 microns, specifically, such as 30 microns, 25 microns, 20 microns, etc.
  • the thickness of the isolation film is as small as possible, which is conducive to the rapid conduction of heat between the temperature regulating device 100 and the reaction sample, and reduces the lag time of the reaction sample temperature relative to the temperature of the temperature regulating device 100.
  • the body 1 and/or the sample holding cavity 14 are flat structures. It is understood that the flat structure may mean that the dimension of the body 1 or the sample holding cavity 14 in the thickness direction (the direction in which the first wall 11 and the second wall 12 are arranged is the thickness direction (Y) of the carrier) is smaller than the dimension in the width direction (X) or the length direction (Z). As an example, the ratio of the dimension of the sample holding cavity 14 or the body 1 perpendicular to its thickness direction (Y) to its thickness direction (Y) is greater than 5:1, such as a dimension ratio of 50:1 to 100:1.
  • the sample holding cavity 14 is a cuboid, and the ratio of the length to the thickness of the cuboid can be greater than 5:1, such as a ratio of 50:1 to 100:1.
  • the ratio of the dimensions is 90:1.
  • the dimension of the sample holding cavity 14 in the thickness direction can be 0.3-1.0 mm, and the width and length of the sample holding cavity 14 are about 10 mm and 20 mm, respectively.
  • the sample holding cavity 14 can also be a cylindrical structure, and the ratio of the diameter to the thickness is greater than 5:1, such as a thickness of 0.3-1.0 mm and a diameter of 5-20 mm.
  • the cross-section of the sample holding cavity 14 can be a polygon or an ellipse, etc.
  • the cross-section of the sample holding cavity 14 can be a polygon or an ellipse, etc.
  • the reaction sample in the flat structure body 1 or sample holding cavity 14 is thin, the center of the reaction sample is very close to the surface of the liquid, the temperature of the reaction sample can reach a uniform temperature in a very short time, and the flat structure can make the contact area between the reaction sample and the temperature adjustment device 100 large, the heat transfer efficiency is high, so that the reaction sample temperature rise and fall speed and detection efficiency are greatly improved.
  • the inner diameter of the tube carrier is relatively large compared to the flat structure sample holding cavity 14, the center of the reaction sample is very close to the surface of the liquid, the temperature of the reaction sample needs a long time to reach a uniform temperature, the reaction sample temperature rise and fall speed is low, and the detection efficiency is low.
  • the flat structure carrier or sample holding cavity is vertically arranged, that is, as shown in FIG. 1e and FIG. 2e, the thickness direction (Y) of the carrier is parallel to the vertical direction (Z).
  • the gas released by the reaction sample in the sample holding chamber 14 during the heating process in the amplification stage is more likely to float to the upper end of the main body 1 instead of being dispersed everywhere in the sample holding chamber 14.
  • the carrier is placed vertically, and thermal convection occurs in the liquid, which is conducive to further mixing the liquid and improving the temperature uniformity of the reaction sample.
  • the first wall 11 and the second wall 12 are both deformable walls at least at the positions opposite to the sample holding chamber 14, and the temperature regulating device 100 heats and cools on both sides of the carrier (i.e., the first wall 11 and the second wall 12), further improving the heating and cooling speed of the reaction sample.
  • the deformable wall can be sealed and connected to the side wall 13 by heat pressing, laser welding or ultrasonic welding.
  • the side wall 13 is made of a transparent material, and the fluorescence detection device 200 can perform fluorescence detection on the reaction sample through the transparent side wall 13.
  • the fluorescence detection device 200 can perform fluorescence detection on the reaction sample through the transparent side wall 13.
  • the reaction sample can be directly detected after amplification without transferring the carrier and the reaction sample, thereby improving the detection efficiency.
  • the position of the first wall 11 (or the second wall 12) opposite to the sample holding cavity 14 is a deformable wall, that is, the first wall 11 of the carrier is deformable, and the second wall 12 is not deformable, or the deformation degree is very small relative to the first wall 11.
  • the thickness of the second wall 12 is not greater than 0.5 mm, so as to reduce the amount of heat absorbed by the second wall 12 during the heating and cooling process, thereby reducing the time for the reaction sample temperature to reach equilibrium and accelerating the heating and cooling speed of the reaction sample.
  • the second wall 12 and/or the side wall 13 are made of a transparent material, and fluorescence detection of the reaction sample can be performed through the second wall 12 or the side wall 13.
  • fluorescence detection is performed through the second wall 12
  • the temperature adjustment device 100, the carrier and the fluorescence detection device 200 can be arranged in sequence, so that the device is more compact and occupies less space.
  • the second wall 12 and the side wall 13 can also be integrally formed by injection molding to improve the processing efficiency of the carrier. Furthermore, the deformable wall can be sealed and connected to the side wall 13 by heat pressing, laser welding or ultrasonic welding.
  • the transparent material can be dimethylsiloxane (ie PDMS), polypropylene (ie PP), plexiglass (ie PMMA) or polycarbonate (ie PC).
  • PDMS dimethylsiloxane
  • PP polypropylene
  • PMMA plexiglass
  • PC polycarbonate
  • PMMA, PDMS, PP and PC are optically transparent materials with good biocompatibility, so they can meet the needs of fluorescence detection and have no effect on the reaction samples.
  • the body 1 may also be provided with a liquid inlet channel 3, through which the sample holding chamber 14 and the sealing chamber 15 are connected, and then liquid is injected and exhausted through the sealing chamber 15 and the liquid inlet channel 3. At this time, after the sealing member 2 is sealed and inserted into the sealing chamber 15, the air in the sealing chamber 15 and the liquid inlet channel 3 is compressed.
  • the cross-sectional area of the liquid inlet channel 3 is preferably smaller than the cross-sectional area of the sealing chamber 15, thereby increasing the pressure difference between the body 1 and the outside.
  • the sample holding chamber 14 and the liquid inlet channel 3 can be formed by cutting a groove on the second wall 12 and then covering the first wall 11 on one side of the groove on the second wall 12, such as forming the groove integrally by injection molding or forming the groove by machining.
  • the carrier when the reaction sample is amplified, the carrier is placed vertically, the sealed cavity 15 is located at the upper end of the carrier, the upper end of the liquid inlet channel 3 is connected to the sealed cavity 15, and the lower end of the liquid inlet channel 3 is connected to the lower end of the sample holding cavity 14. Therefore, when liquid is injected into the sample holding cavity 14, the reaction sample enters from the lower end of the sample holding cavity 14, and the air is located above the reaction sample. The air floats up, making it easier to discharge the air, such as through the liquid inlet channel 3.
  • the main body 1 is optionally provided with an exhaust channel 4, one end of the exhaust channel 4 is connected to the sealing cavity 15, and the other end is connected to the sample holding cavity 14. At this time, when the liquid inlet channel 3 is filled with liquid, the air in the sample holding cavity 14 can be exhausted through the exhaust channel 4, avoiding the sample holding cavity 14. Air remains in the nanocavity 14, thereby ensuring that the reaction sample is quickly heated and cooled.
  • the lower end of the exhaust channel 4 is connected to the upper end of the sample holding chamber 14, and the upper end is connected to the sealing chamber 15.
  • the air floats up into the lower end of the exhaust channel 4, and then is exhausted from the exhaust channel 4.
  • the carrier is placed vertically. Before the sample holding chamber 14 is filled with the reaction sample, the lower end of the exhaust channel 4 will not be blocked by the reaction solution, and the exhaust channel 4 can be exhausted all the time.
  • a liquid injection hole 5 is provided between the liquid inlet channel 3 and the sealed cavity 15 , and a liquid transfer tool such as a pipette tip can be inserted into the liquid injection hole 5 to inject liquid into the sample receiving cavity 14 .
  • the inner wall of the injection hole 5 is a truncated cone structure, the large diameter end of the injection hole 5 is connected to the sealing cavity 15 , and the small diameter end is connected to the liquid inlet channel 3 , so as to adapt to the pipette tip for pipetting operations.
  • the seal 2 and the sealing cavity 15 are interference-fitted, which can seal the sealing cavity 15 on the one hand, and on the other hand, can prevent the seal 2 from being opened when there is a pressure difference between the inside of the body 1 and the outside.
  • the seal 2 and the sealing cavity 15 can also be sealed and connected in other ways.
  • the sealing member 2 includes a first plug (not shown in the figure), which is sealed and arranged in the sealing cavity 15. At this time, the first plug is interference-fitted with the sample holding cavity 14, and the volume of the first plug inserted into the sealing cavity 15 is the volume of the compressed air in the body 1.
  • the first plug is made of elastic material such as rubber or silicone.
  • the sealing member 2 is of the above structure, since the first plug is interference-fitted with the sample holding cavity 14 , the operator needs to exert a relatively large force to insert the first plug into the sealing cavity 15 .
  • the sealing member 2 includes a second plug 21 and a sealing portion 22 connected to each other.
  • the sealing portion 22 is arranged around the outer periphery of the second plug 21 and is spaced from the outer periphery of the second plug 21.
  • the outer periphery of the sealing portion 22 can be in sealing contact with the cavity wall of the sealing cavity 15.
  • the second plug 21 and the sealing portion 22 are both inserted into the sealing cavity 15, and the volume of the second plug 21 and the sealing portion 22 inserted into the sealing cavity 15 is the volume of the compressed air in the body 1.
  • the sealing portion 22 may be interference fit with the sealing cavity 15.
  • the sealing portion 22 is subjected to the force of the cavity wall of the sealing cavity 15, so that the sealing cavity 15 moves toward the second plug 21, thereby reducing the friction between the cavity wall of the sealing cavity 15 and the sealing portion 22, thereby facilitating the insertion of the sealing member 2 into the sealing cavity 15.
  • the sealing portion 22 is made of elastic material such as rubber or silicone.
  • a sealing convex ring 221 is formed on the outer periphery of the sealing part 22, and the sealing convex ring 221 is in sealing contact with the cavity wall of the sealed cavity 15.
  • the sealing convex ring 221 can seal the sealed cavity 15, and at the same time, can reduce the contact area between the sealing part 22 and the cavity wall of the sealed cavity 15, thereby reducing the friction between the cavity wall of the sealed cavity 15 and the sealing part 22.
  • the volume of the seal 2 inserted into the sealing cavity 15 determines the pressure difference between the inside and outside of the carrier.
  • the seal 2 optionally further includes a positioning cap 23.
  • the ends of the second plug 21 and the sealing portion 22 are both connected to the positioning cap 23, and the positioning cap 23 can abut against the outer surface of the body 1. That is, the positioning cap 23 plays a positioning role. When the positioning cap 23 abuts against the outer surface of the body 1, the seal 2 is inserted into place.
  • the thickness of the carrier is small.
  • the body 1 also includes an annular wall 16, which is connected to the side wall 13.
  • the space inside the annular wall 16 forms the sealed cavity 15.
  • the annular wall 16 is a circular ring structure, and the cross section of the sealed cavity 15 is a cylindrical structure.
  • the first plug is a cylindrical structure, and the cross section of the sealing portion 22 is a circular ring, thereby sealing the sealed cavity 15.
  • the annular wall 16 may also be a polygonal ring such as a square ring, as long as the annular wall 16 extends continuously and forms a circumferentially closed structure.
  • the seal 2 can be connected to the body 1 via a connecting strip 24 .
  • the seal 2 and the body 1 may be integrally formed to facilitate processing of the carrier.
  • the thickness of the first wall 11 and the second wall 12 are both relatively thin, so as to improve the heat conduction efficiency between the reaction sample and the temperature regulating device 100, and reduce the heat loss caused by the heat conducted to the first wall 11 and the second wall 12. Therefore, in order to meet the requirements of rapid temperature rise and fall of the reaction sample and sufficient strength of the carrier, as shown in FIG. 9 , the carrier may optionally further include a reinforcement portion, which is used to strengthen the main body 1.
  • the reinforcing portion includes a first reinforcing portion 7, which can be a strip structure to improve the strength of the carrier and prevent the carrier from twisting and deforming.
  • the first reinforcing portion 7 is far away from the sample holding cavity, which will not affect the rapid heating and cooling of the reaction sample and the detection of the reaction sample.
  • the thickness of the first reinforcing portion 7 is greater than the thickness of the body 1 (that is, the size of the first reinforcing portion 7 in the thickness direction (Y) is greater than the size of the body 1 in the thickness direction (Y)) to improve the strength of the carrier and prevent the carrier from twisting and deforming.
  • the body 1 is a rectangular parallelepiped structure, and the first reinforcing portion 7 is arranged on one or both sides of the width direction (X) of the body 1.
  • the width direction (X), the thickness direction (Y) and the vertical direction (Z) are perpendicular to each other.
  • the reinforcement part also includes a second reinforcement part 8, and the second reinforcement part 8 is disposed on one or both sides of the length direction (i.e., the vertical direction (Z)) of the body 1, and the thickness of the second reinforcement part 8 is greater than the thickness of the body 1, so as to improve the strength of the carrier and prevent the carrier from being twisted and deformed.
  • the second reinforcement part 8 is disposed at the upper end of the body 1, and optionally, the second reinforcement part 8 is located between the two first reinforcement parts 7, and further, the second reinforcement part 8 is disposed close to the upper ends of the two first reinforcement parts 7.
  • the outer surface of the second wall 12 can be connected to the reinforcing rib 6 , so as to strengthen the carrier, prevent deformation of the carrier such as twisting, and reduce expansion caused by pressure difference.
  • the carrier In order to insert the carrier between two temperature regulating devices 100 (as shown in FIG. 1e ) or between the temperature regulating device 100 and the fluorescence detection device 200 (as shown in FIG. 2e ), the carrier also includes a handle 10 , which is connected to one end of the body 1 .
  • the handle 10 is a strip structure.
  • the carrier may further include a positioning portion 9 for positioning the carrier.
  • the positioning portion 9 is a positioning notch provided on the body 1 or the reinforcement portion, and illustratively, the positioning notch is provided on the first reinforcement portion 7.
  • a positioning protrusion 105 may be provided on the temperature adjustment device 100, such as connecting the positioning protrusion 105 on the card slot 104, inserting the carrier into the card slot 104, and when the positioning protrusion 105 enters the positioning notch, it indicates that the carrier is inserted into the preset position.
  • the carrier provided in this embodiment adopts a deformable membrane, so that the temperature regulating device 100 can quickly transfer heat to the reaction sample in the sample holding cavity 14 through the deformable membrane; by reasonably designing the pressure inside the carrier, the internal air pressure is relatively large, and the deformable membrane can form a good thermal contact with the temperature regulating device 100, thereby avoiding the air gap between the deformable membrane and the temperature regulating device 100 affecting heat conduction; the sample holding cavity 14 adopts a flat structure design, and the reaction sample temperature can quickly reach equilibrium; through the above technical means, the reaction sample temperature can respond quickly, reducing the lag time between the reaction sample temperature and the temperature of the temperature regulating device 100, thereby reducing the reaction time of PCR.

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Abstract

La présente invention concerne un support, comportant un corps et un élément d'étanchéité, le corps étant pourvu d'une cavité de maintien de l'échantillon et d'une cavité d'étanchéité, la cavité de maintien de l'échantillon étant en communication avec la cavité d'étanchéité ; une paroi de la cavité de maintien de l'échantillon est une paroi déformable, et au moins une partie de l'élément d'étanchéité peut être insérée dans la cavité d'étanchéité de manière étanche, de manière à au moins comprimer le gaz dans la cavité d'étanchéité et à permettre à la paroi déformable de faire saillie vers l'extérieur pour permettre à la paroi déformable d'être en contact étroit avec un dispositif de régulation de la température. Le support peut accélérer les vitesses de chauffage et de refroidissement d'un échantillon de réaction et améliorer l'efficacité de l'amplification.
PCT/CN2024/105017 2023-07-11 2024-07-11 Support Pending WO2025011622A1 (fr)

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CN202310852944.0A CN119307344A (zh) 2023-07-11 2023-07-11 一种载体
CN202310852944.0 2023-07-11

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Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005001434A2 (fr) * 2003-06-23 2005-01-06 Applera Corporation Bouchons pour puits a echantillons et microcartes destinees a recevoir des materiels biologiques
JP2009139138A (ja) * 2007-12-04 2009-06-25 Toppan Printing Co Ltd 反応容器
WO2010060509A1 (fr) * 2008-11-03 2010-06-03 Zenteris Gmbh Cartouche et dispositif pour analyser des échantillons biologiques par des réactions biologiques à température contrôlée
JP2018023334A (ja) * 2016-08-12 2018-02-15 セイコーエプソン株式会社 熱サイクル装置、反応容器、および核酸増幅方法
CN112041073A (zh) * 2018-03-23 2020-12-04 株式会社百奥尼 高速聚合酶链式反应分析板
CN113164955A (zh) * 2018-11-29 2021-07-23 康特姆斯集团有限公司 改进的微流体设备、系统和方法
CN216972496U (zh) * 2022-03-23 2022-07-15 无锡百泰克生物技术有限公司 一种单份快速pcr反应芯片
CN115386468A (zh) * 2022-08-10 2022-11-25 上海妙灵生物工程有限公司 一种异型生化试管
WO2023072090A1 (fr) * 2021-10-25 2023-05-04 上海透景生命科技股份有限公司 Ensemble tube de réaction en chaîne de la polymérase

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005001434A2 (fr) * 2003-06-23 2005-01-06 Applera Corporation Bouchons pour puits a echantillons et microcartes destinees a recevoir des materiels biologiques
JP2009139138A (ja) * 2007-12-04 2009-06-25 Toppan Printing Co Ltd 反応容器
WO2010060509A1 (fr) * 2008-11-03 2010-06-03 Zenteris Gmbh Cartouche et dispositif pour analyser des échantillons biologiques par des réactions biologiques à température contrôlée
JP2018023334A (ja) * 2016-08-12 2018-02-15 セイコーエプソン株式会社 熱サイクル装置、反応容器、および核酸増幅方法
CN112041073A (zh) * 2018-03-23 2020-12-04 株式会社百奥尼 高速聚合酶链式反应分析板
CN113164955A (zh) * 2018-11-29 2021-07-23 康特姆斯集团有限公司 改进的微流体设备、系统和方法
WO2023072090A1 (fr) * 2021-10-25 2023-05-04 上海透景生命科技股份有限公司 Ensemble tube de réaction en chaîne de la polymérase
CN216972496U (zh) * 2022-03-23 2022-07-15 无锡百泰克生物技术有限公司 一种单份快速pcr反应芯片
CN115386468A (zh) * 2022-08-10 2022-11-25 上海妙灵生物工程有限公司 一种异型生化试管

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