A liquid nucleic acid sample volume measurement consumable device and method

Abstract

The present application relates to the technical field of nucleic acid sample preparation and amplification detection, and specifically to a liquid-phase nucleic acid sample volume metering consumable device and method, comprising a sample collection tube, a metering component connected to the sample collection tube, a sealing component arranged between the sample collection tube and the metering component, a reaction tube connected to the metering component, and a cut-off component arranged in a cavity of the consumable device, wherein the metering component comprises a metering shell connected to the sealing component, a metering tube connected to the cut-off component through an upper opening, and an opening and closing component that blocks the lower end of the metering tube, the sample collection tube is provided with an external connection structure on the outer side of one open end, the sample collection tube is connected to the sealing component through the external connection structure, and a fixing component for fixing the reaction tube is arranged on the outer periphery of the metering shell, thereby realizing the purpose of integrating, automating and miniaturizing multi-step operations in the nucleic acid sample preparation process.

Description

Technical Field

[0001] The present application relates to the technical field of nucleic acid samples and amplification detection, and in particular to a liquid-phase nucleic acid sample volume measurement consumable device and method. Background Art

[0002] Sample preparation for nucleic acid testing involves multiple steps. These steps include releasing nucleic acids from tissues, cells or microorganisms, removing impurities or inhibitors, concentrating nucleic acids, determining sample volume, mixing a certain volume of sample with reagents, and transferring the mixture to a reaction tube for amplification reaction.

[0003] In the prior art, there are two main methods for sample preparation for nucleic acid testing, namely manual operation and automated operation. The manual operation method is that the experimenter completes the operation in the laboratory by using instruments such as centrifuges, pipettes, and vortexers; the automated method is an all-in-one machine controlled by a computer to automatically complete all operation steps. Manual operation is labor-intensive, cumbersome, and prone to errors; automated operation involves high-cost instrument systems, and the instrument systems occupy a large laboratory space. Automated instrument systems are good for high-throughput operations, but they are cost-effective for low-throughput operations and point-of-care diagnosis (POCT). For this reason, a liquid-phase nucleic acid sample volume metering consumables device and method are needed. Summary of the invention

[0004] In view of the shortcomings of the prior art, the purpose of this application is to provide a liquid-phase nucleic acid sample volume metering consumables device and method to solve the technical problems in the prior art of high labor intensity in manual operation during nucleic acid sample preparation and low cost-effectiveness of automated detection.

[0005] The above-mentioned purpose of the present application is achieved through the following technical solutions: a liquid-phase nucleic acid sample volume metering consumable device, comprising a sample collection tube, a metering component connected to the sample collection tube, a sealing component arranged between the sample collection tube and the metering component, a reaction tube connected to the metering component, and a cut-off component arranged in the cavity of the consumable device, the metering component comprising a metering shell connected to the sealing component, a metering tube communicating with the cut-off component, and an opening and closing component located at the lower end of the metering tube, the opening at the lower end of the metering tube is blocked by the opening and closing component, the sample collection tube is provided with an opening, an external connection structure is provided on the outer side of one end of the sample collection tube provided with the opening, the sample collection tube is connected to the sealing component through the external connection structure, and a fixing component for fixing the reaction tube is provided on the outer periphery of the metering shell.

[0006] By adopting the above technical solution, the sample collection tube is functionally coupled with an ultrasonic generator and / or a heater to process the liquid sample in the sample collection tube. The ultrasonic generator is used for cell disruption and nucleic acid release. The function of the heater is to raise the temperature of the liquid in the sample collection tube to the required temperature, which is 80 o C-110 o C, such as 90 o C or 95 o C or 98 o C or 100 o C or 105 o C. The heater can be a semiconductor heater such as a Peltier heater or a resistance heater or an infrared heater or an electromagnetic induction heater or a microwave heater. The nucleic acid liquid sample is placed in the sample collection tube, and then the sample collection tube is connected to the sealing component through the external connection structure, so that the sealing component is located between the metering component and the sample collection tube, and the gap between the sample collection tube and the metering component is sealed, and then the heater is started to generate a sufficiently high temperature in the sample collection tube to lyse eukaryotic cells or prokaryotic cells or viruses, etc., and release nucleic acids. The temperature can also be high enough to denature enzymes and inactivate pathogens. In addition, the increased temperature can generate sufficient internal pressure, which is sufficient to press the nucleic acid liquid sample into the metering tube, and the nucleic acid liquid sample flows into the metering tube through the sealing component and the cut-off component. After the nucleic acid liquid sample overflows in the metering tube, it will trigger the cut-off component, so that the cut-off component closes the passage between the sample collection tube and the metering component, so that the nucleic acid liquid sample no longer flows into the metering tube, so that a quantitative nucleic acid liquid sample is maintained in the metering tube. The staff opens the passage between the metering tube and the reaction tube through the opening and closing component, so that a quantitative nucleic acid liquid sample flows into the reaction tube. Liquid samples can flow into the reaction tube by gravity or centrifugal force, and centrifugal force can be generated manually or by a centrifuge. This allows staff to easily complete nucleic acid sample preparation, reducing preparation costs while making nucleic acid sample preparation more convenient, and achieving the goal of integrating multiple steps in the nucleic acid sample preparation process, automation, and miniaturization.

[0007] Furthermore, the sealing assembly includes a sealing shell and sealing gaskets fixedly arranged at the upper and lower ends of the sealing shell. An internal connecting structure is provided inside the sealing shell at one end close to the sample collection tube. The external connecting structure of the sample collection tube is connected to the internal connecting structure of the sealing shell. The external connecting structure and the internal connecting structure are threads or snaps.

[0008] By adopting the above technical solution, the sample collection tube is twisted and connected to the sealing shell through the external connection structure, so that the sample collection tube extends into the sealing shell, and the end face of the sample collection tube presses the sealing gasket, thereby preventing the nucleic acid liquid sample from flowing out.

[0009] Furthermore, a filter pad is provided in the sealing component, and the filter pad has the performance of filtering insoluble particles or adsorbing soluble non-nucleic acid inorganic molecules or organic molecules. The filter pad is membrane-shaped or pad-shaped, and its matrix is ​​a mesh material or a porous granular material.

[0010] By adopting the above technical solution, the filter pad can completely or partially remove impurities in the nucleic acid liquid sample, wherein the impurities are insoluble particles or inorganic or organic molecules other than nucleic acids, and the matrix material of the filter pad includes cellulose, nylon, resin, glass, metal, metal oxide or a combination thereof, which can be formed into a mesh or porous granular form. The surface of these matrices can be functionally modified, such as carboxyl, amino, sulfonic, hydroxyl, benzyl, fatty, etc.

[0011] Furthermore, the truncation assembly includes a support plate, a truncation plate located below the support plate, a guide tube used to connect the truncation plate and extend below the truncation plate into the metering tube, and a truncation block arranged below the truncation plate. The truncation plate and the guide tube are made of elastic material, and a through opening for the flow of samples is opened on the support plate.

[0012] Furthermore, the support plate is disposed in a cavity of the metering component, or the support plate is disposed in a cavity of the sealing component.

[0013] Furthermore, the truncated block is a water-swellable polymer, and the water-swellable polymer includes a hydrogel.

[0014] By adopting the above technical scheme, the nucleic acid liquid sample flows to the support plate through the sealing shell of the sealing component or the metering shell of the metering component, then flows to the cut-off plate through the opening on the support plate, then flows into the guide tube through the cut-off plate, and flows into the metering tube under the guidance of the guide tube. When the nucleic acid liquid sample in the metering tube is overflowing, the overflowing nucleic acid liquid sample will flow into the placement space and be absorbed by the cut-off block. After the cut-off block absorbs the nucleic acid liquid sample, the cut-off block excites and squeezes the guide tube, so that the guide tube is closed, the passage between the metering component and the sample collection tube is cut off, and the nucleic acid liquid sample inside the metering component is quantified, wherein the elastic material of the cut-off plate and the guide tube is a thermoplastic polyurethane elastomer material, a silicone material, a high-temperature nylon material or a carbon nanotube-containing material, and any water-soluble or hydrophilic polymer can form a hydrogel through certain chemical crosslinking or physical crosslinking. These polymers can be divided into two categories: natural and synthetic according to their sources. Natural hydrophilic polymers include polysaccharides (starch, cellulose, sea acid, hyaluronic acid, chitosan, etc.) and polypeptides (collagen, poly L-lysine, poly L-glutamic acid, etc.); synthetic hydrophilic polymers include alcohol, acrylic acid and its derivatives (polyacrylic acid, polymethacrylic acid, polyacrylamide, poly N-polysubstituted acrylamide, etc.).

[0015] Furthermore, the fixing assembly includes a fixing buckle fixedly arranged on the outer periphery of the metering shell and a clamping piece fixedly arranged on the periphery of the upper end of the reaction tube.

[0016] By adopting the above technical solution, the reaction tube is pushed to cover the metering tube, so that the clamping piece is clamped by the fixing buckle, so that the reaction tube is fixed.

[0017] Furthermore, the opening and closing assembly includes a twistable cover installed on one end of the metering tube close to the reaction tube and a stopper sleeved on the twistable cover, the reaction tube and the stopper have no sliding fit, and the metering tube and the metering shell have no sliding fit.

[0018] By adopting the above technical solution, after the reaction tube covers the metering tube, the reaction tube is rotated so that the reaction tube drives the stopper to rotate, and the twist-off cover is twisted off. Under the action of gravity or centrifugal force, the liquid sample of the preset volume in the metering tube is transferred to the reaction tube, and the metering tube and the twist-off cover of the opening and closing component can be injection molded at one time, so that the metering tube is connected to the reaction tube when the twist-off cover is separated from the metering tube. The internal space of each component of the metering consumable device is sealed and isolated from the outside, that is, fully sealed; wherein the metering component and the reaction tube can rotate with the circumference without destroying the sealing structure.

[0019] Furthermore, a current limiting device is provided above the cutoff component, and the current limiting device is a heat-sensitive structure or a pressure-sensitive structure.

[0020] By adopting the above technical solution, in order to prevent the sample from flowing into the metering tube too early, a flow limiting device is provided above the cut-off component, such as a heat-sensitive structure or a pressure-sensitive structure. The pressure-sensitive structure can be a pressure-sensitive membrane; its function is that when the pressure difference on both sides of the pressure-sensitive membrane reaches a preset pressure, the device can be turned on. The structure can be a plastic film that breaks at a given temperature or at a given air pressure; this structure can be turned on after the liquid sample reaches a predetermined temperature, such as 90 o C or 95 o C or 100 o C or 105 o C allows liquid to flow through; or when the air pressure reaches above 1.5 atmospheres, above 2.0 atmospheres, above 2.5 atmospheres, or above 3 atmospheres, liquid can flow through.

[0021] A liquid-phase nucleic acid sample volume measurement method is applied to the above-mentioned liquid-phase nucleic acid sample volume measurement consumable device, comprising the following steps:

[0022] S1. Connecting a sample collection tube containing a nucleic acid sample liquid to be detected to a sealing component to form a fully sealed nucleic acid sample volume metering consumable device;

[0023] S2, coupling a heater to the sample collection tube and starting the heater, the heater can inactivate cells or viruses and release nucleic acid molecules;

[0024] S3. The liquid sample in the sample tube is heated to generate a relatively high air pressure. Under this high air pressure, the lower liquid is pushed into the metering tube through the filter pad, the cut-off plate and the guide tube;

[0025] S4, when the liquid sample overflows in the metering tube, the cut-off block absorbs the overflowing liquid sample and expands, thereby pushing the cut-off plate and closing the through hole and / or the guide tube of the support plate, thereby cutting off the passage of the liquid from the sample tube to the metering tube;

[0026] S5, rotating the reaction tube so that the reaction tube drives the stopper to rotate, twisting off the twist-off cover, and opening the passage between the metering tube and the reaction tube;

[0027] S6. Under the action of gravity or centrifugal force, the preset volume of liquid sample in the measuring tube is transferred to the reaction tube.

[0028] In summary, the present application includes at least one of the following beneficial technical effects:

[0029] Through the setting of the liquid-phase nucleic acid sample volume metering consumable device, the nucleic acid liquid sample is poured into the sample collection tube, and then the sample collection tube is connected to the sealing component through the external connection structure, so that the sealing component is located between the metering component and the sample collection tube, and the gap between the sample collection tube and the metering component is sealed, and then the heater is started to generate a sufficiently high temperature in the sample collection tube to lyse eukaryotic cells or prokaryotic cells or viruses, etc., and release nucleic acids. The temperature can also be high enough to denature enzymes and inactivate pathogens. In addition, the increased temperature can generate sufficient internal pressure, which is sufficient to press the nucleic acid liquid sample into the metering tube. The nucleic acid liquid sample flows into the metering tube through the sealing component and the cut-off component. After the nucleic acid liquid sample overflows in the metering tube, the cut-off component is triggered, so that the cut-off component closes the passage between the sample collection tube and the metering component, so that the nucleic acid liquid sample no longer flows into the metering tube, so that a quantitative nucleic acid liquid sample is maintained in the metering tube. The staff opens the passage between the metering tube and the reaction tube through the opening and closing component, so that a quantitative nucleic acid liquid sample flows into the reaction tube. The liquid sample flows into the reaction tube through gravity or centrifugal force, and the centrifugal force can be generated manually or with the help of a centrifuge. This allows staff to easily complete nucleic acid sample preparation, reduces preparation costs and makes nucleic acid sample preparation more convenient, achieving the goal of integrating, automating and miniaturizing multiple steps in the nucleic acid sample preparation process. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is an exploded schematic diagram of the overall structure of Example 1;

[0031] Figure 2 is the overall structure of the uncoupled heater in Example 1 and its cross-sectional view;

[0032] Figure 3 is a cross-sectional view of the overall structure of the uncoupled heater in Example 2;

[0033] Figure 4 yes Figure 3 Enlarged view of part A in the middle.

[0034] Figure numerals: 1. sample collection tube; 2. metering component; 20. metering shell; 7. cut-off component; 70. cut-off plate; 71. guide tube; 72. cut-off block; 73. support plate; 22. metering tube; 23. opening and closing component; 230. twist-off cover; 231. stopper; 3. sealing component; 30. sealing shell; 31. sealing gasket; 32. filter pad; 4. heater; 5. fixing component; 50. fixing buckle; 51. snap-on piece; 6. reaction tube; 8. current limiting device. DETAILED DESCRIPTION

[0035] The present application is further described in detail below in conjunction with the accompanying drawings.

[0036] Example 1, reference Figure 1 as well as Figure 2 , including a sample collection tube 1, a metering component 2 connected to the sample collection tube 1, a sealing component 3 arranged between the sample collection tube 1 and the metering component 2, and a reaction tube 6 connected to the metering component 2, the metering component 2 includes a metering shell 20 connected to the sealing component 3, a truncation component 7 arranged in the metering shell 20 of the metering component 2, a metering tube 22 connected to the truncation component 7, and an opening and closing component 23 located at the lower end of the metering tube 22, the opening at the lower end of the metering tube 22 is blocked by the opening and closing component 23, the sample collection tube 1 is provided with an opening, an external connection structure is provided on the outer side of one end of the sample collection tube 1 provided with the opening, the sample collection tube 1 is connected to the sealing component 3 through the external connection structure, a fixing component 5 for fixing the reaction tube 6 is provided on the outer periphery of the metering shell 20, the truncation component 7 includes a support plate 73 and a truncation block 72, a heater 4 and / or an ultrasonic generator are functionally coupled to the sample collection tube 1 to process the liquid sample in the sample collection tube. The ultrasonic generator is used for cell disruption and nucleic acid release. The function of the heater 4 is to raise the temperature of the liquid in the sample collection tube 1 to the required temperature, the required temperature is 80 o C-110 o C can be 90 o C or 95 o C or 100 o C or 105 oC. The heater 4 is a semiconductor heater or a resistance heater or an infrared heater or an electromagnetic heater or a microwave heater. The nucleic acid liquid sample is placed in the sample collection tube 1, and then the sample collection tube 1 is connected to the sealing component 3 through the external connection structure, so that the sealing component 3 is located between the metering component 2 and the sample collection tube 1, and the gap between the sample collection tube 1 and the metering component 2 is sealed. Then the heater 4 is started to generate a sufficiently high temperature in the sample collection tube 1 to lyse eukaryotic cells or prokaryotic cells or viruses, etc., and release nucleic acids. The temperature can also be high enough to denature enzymes and inactivate pathogens. In addition, the increased temperature can generate sufficient internal pressure, which is sufficient to press the nucleic acid liquid sample into the metering tube 22. The nucleic acid liquid sample flows into the metering tube 22 through the sealing component 3 and the cut-off component 7. After the nucleic acid liquid sample overflows in the metering tube 22, it will trigger the cut-off component 7, so that the cut-off component 7 closes the passage between the sample collection tube 1 and the metering component 2, so that the nucleic acid liquid sample no longer flows into the metering tube 22, so that a quantitative nucleic acid liquid sample is maintained in the metering tube 22. The staff opens the passage between the metering tube 22 and the reaction tube 6 through the opening and closing component 23, so that a quantitative nucleic acid liquid sample flows into the reaction tube 6. The liquid sample can flow into the reaction tube 6 by gravity or centrifugal force, and the centrifugal force can be generated manually or by a centrifuge. In this way, the staff can easily complete the preparation of nucleic acid samples, reduce the preparation cost, make the preparation of nucleic acid samples more convenient, and achieve the purpose of integrating multiple steps of the nucleic acid sample preparation process, automating and miniaturizing.

[0037] In this embodiment, the sealing assembly 3 includes a sealing shell 30 and sealing gaskets 31 fixedly arranged at the upper and lower ends of the sealing shell 30. An internal connecting structure is provided inside the sealing shell 30 at one end close to the sample collection tube 1. The external connecting structure of the sample collection tube 1 is connected to the internal connecting structure of the sealing shell 30. The external connecting structure and the internal connecting structure are threads or buckles. By twisting the sample collection tube 1 to connect to the sealing shell 30 through the external connecting structure, the sample collection tube 1 is extended into the sealing shell 30, so that the end face of the sample collection tube 1 presses the sealing gasket 31 to prevent the nucleic acid liquid sample from flowing out.

[0038] In this embodiment, a filter pad 32 is provided in the sealing component 3. The filter pad 32 has the performance of filtering insoluble particles or adsorbing soluble non-nucleic acid inorganic molecules or organic molecules. The filter pad 32 is in the form of a membrane or a pad, and its matrix is ​​a mesh material or a porous granular material. The filter pad 32 can remove all or part of the impurities in the nucleic acid liquid sample. The impurities are insoluble particles or non-nucleic acid inorganic molecules or organic molecules. The matrix material of the filter pad 32 includes cellulose, nylon, resin, glass, metal, metal oxide or a combination thereof, which can form a mesh or porous granular shape. The surface of these matrices can be functionally modified, such as carboxyl, amino, sulfonic, hydroxyl, benzyl, fatty, etc.

[0039] In this embodiment, a flow limiting device 8 is provided above the cut-off component. The flow limiting device 8 may be a heat-sensitive structure or a pressure-sensitive structure. The structure may be a plastic film that breaks at a predetermined temperature or at a predetermined pressure. When the liquid sample reaches a predetermined temperature, such as 90 o C or 95 o C or 100 o C or 105 o C allows the liquid to flow through; or when the air pressure reaches 1.5 atmospheres or above, 2.0 atmospheres or above, 2.5 atmospheres or above, or 3 atmospheres or above, the liquid can be allowed to flow through, which can effectively prevent the sample from flowing into the metering tube 22 too early.

[0040] In this embodiment, the truncation assembly 7 includes a support plate 73, a truncation plate 70 located below the support plate 73, a guide tube 71 for connecting the truncation plate 70 and extending below the truncation plate 70 into the metering tube 22, and a truncation block 72 arranged below the truncation plate 70. The truncation plate 70 and the guide tube 71 are made of elastic material. A through hole for the sample to flow is opened on the support plate 73. The support plate 73 is arranged in the sealing assembly 3. The truncation block 72 is used to push the truncation plate 70 to move upward. The truncation plate 70 and the metering tube 22 are connected. 22, a space is left between them to form a placement space, the cut-off block 72 is located in the placement space and surrounds the guide tube 71, the cut-off plate 70 is connected to the guide tube 71 as a whole, the cut-off block 72 is a water-swelling polymer, and the water-swelling polymer includes a hydrogel, wherein the elastic material of the cut-off plate 70 and the guide tube 71 is a thermoplastic polyurethane elastomer material, a silicone material, a high-temperature nylon material or a carbon nanotube-containing material, and any water-soluble or hydrophilic polymer can form a hydrogel through certain chemical crosslinking or physical crosslinking. These polymers can be divided into two categories according to their sources: natural and synthetic. Natural hydrophilic polymers include polysaccharides (starch, cellulose, alginic acid, hyaluronic acid, chitosan, etc.) and polypeptides (collagen, poly-L-lysine, poly-L-glutamic acid, etc.); synthetic hydrophilic polymers include alcohols, acrylic acid and its derivatives (polyacrylic acid, polymethacrylic acid, polyacrylamide, poly-N-polysubstituted acrylamide, etc.). The nucleic acid liquid sample flows through the sealing shell 30 of the sealing component 3 to the support plate 73, and then flows to the cut-off plate 70 through the opening on the support plate 73, and then flows into the guide tube 71 through the cut-off plate 70, and then flows into the metering tube 22 through the guidance of the guide tube 71. When the nucleic acid specimen sample in the metering tube 22 is overflowing, the overflowing nucleic acid liquid sample will flow into the placement space and be absorbed by the cut-off block 72. After the cut-off block 72 absorbs the nucleic acid liquid sample, the cut-off block 72 excites and squeezes the guide tube 71, so that the guide tube 71 is closed, the passage between the metering component 2 and the sample collection tube 1 is cut off, and the nucleic acid liquid sample inside the metering component 2 is quantified.

[0041] In this embodiment, the fixing assembly 5 includes a fixing buckle 50 fixedly arranged on the outer periphery of the metering shell 20 and a clamping piece 51 fixedly arranged on the periphery of the upper end of the reaction tube 6. By pushing the reaction tube 6 to cover the metering tube 22, the clamping piece 51 is clamped by the fixing buckle 50, so that the reaction tube 6 is fixed.

[0042] In this embodiment, the opening and closing component 23 includes a twist-off cover 230 installed on one end of the metering tube 22 close to the reaction tube 6 and a stopper 231 sleeved on the twist-off cover 230. The reaction tube 6 and the stopper 231 do not slide together, and the metering tube 22 and the metering housing 20 do not slide together. After the reaction tube 6 covers the metering tube 22, the reaction tube 6 is rotated so that the reaction tube 6 drives the stopper 231 to rotate, and the twist-off cover 230 is twisted off. Under the action of gravity or centrifugal force, the liquid sample of the preset volume in the metering tube 22 is transferred to the reaction tube 6. The metering tube 22 and the twist-off cover 230 of the opening and closing component 23 can be injection molded at one time, and the metering tube is connected to the reaction tube 6 when the twist-off cover 230 is separated from the metering tube 22. The internal space of each component of the metering consumable device is sealed and isolated from the outside, that is, fully sealed; wherein the metering component 2 and the reaction tube 6 can rotate with the circumference without destroying the sealing structure.

[0043] Example 2, reference Figure 3 as well as Figure 4 The difference between this embodiment and embodiment 1 is that the support plate 73 of the cut-off component 7 is arranged in the sealed shell 30 of the sealing component 3, the nucleic acid liquid sample flows to the support plate 73 through the sealed shell 30 of the sealing component 3, and then flows to the cut-off plate 70 through the opening on the support plate 73, and then flows into the guide tube 71 through the cut-off plate 70, and then flows into the metering tube 22 under the guidance of the guide tube 71. When the nucleic acid liquid sample in the metering tube 22 is overflowing, the overflowing nucleic acid liquid sample will flow into the placement space and be absorbed by the cut-off block 72. After the cut-off block 72 absorbs the nucleic acid liquid sample, the cut-off block 72 excites and squeezes the guide tube 71, so that the guide tube 71 is closed, the passage between the metering component 2 and the sample collection tube 1 is cut off, and the nucleic acid liquid sample inside the metering component 2 is quantified.

[0044] Specific implementation process: connect the sample collection tube 1 containing the nucleic acid liquid sample to the sealing component 3 to form a fully sealed nucleic acid sample volume metering consumable device, then couple the heater 4 to the sample collection tube 1, and then start the heater 4 to denature the protein molecules in the sample, inactivate the virus and release the nucleic acid. Heating the sample collection tube 1 can also generate a relatively high air pressure for the liquid sample inside the sample collection tube 1. Under this high air pressure, the lower liquid is pushed into the metering tube 22 through the filter pad 32, the cut-off plate 70 and the guide tube 71. The space inside the metering tube 22 is connected to the test tube through the gap of the cut-off component 7. When the liquid enters the metering tube 22 under the action of relatively high pressure and gravity, the air pressure in the upper part of the metering tube 22 will reach a balance with the relatively low pressure in the lower reaction tube 6, so that the liquid flows continuously into the metering tube 22. A heat-sensitive or pressure-sensitive structure is provided between the metering component 2 and the sealing component 3. The structure can be a plastic film that breaks at a given temperature or under a given air pressure. After the liquid sample reaches a predetermined temperature, such as 90 o C or 95 o C or 100 o C or 105 o C allows the liquid to flow through; or when the air pressure reaches 1.5 atmospheres or more, 2.0 atmospheres or more, 2.5 atmospheres or more, or 3 atmospheres or more, the liquid can flow through. When the liquid sample overflows in the metering tube 22, the cut-off block 72 absorbs the overflowing liquid sample and expands, thereby pushing the cut-off plate 70 and closing the through hole or the flow guide tube 71 of the support plate 73, thereby cutting off the passage of the liquid from the sample tube to the metering tube 22, and achieving the purpose of measuring the preset liquid sample volume. This preset liquid sample volume is the volume required to ensure the concentration of the components of the amplification reaction system. Then rotate the reaction tube 6 so that the reaction tube 6 drives the stopper 231 to rotate, twist off the twistable cover 230, and the preset volume of the liquid sample in the metering tube 22 is transferred to the reaction tube 6 under the action of gravity or centrifugal force. The preset liquid sample volume is 1 microliter to 1000 microliters, 10 microliters to 100 microliters, such as 40 microliters, 45 microliters, 50 microliters, 55 microliters, 60 microliters, 65 microliters, and 70 microliters.

[0045] The sample volume measurement of the present application refers to obtaining a preset volume of liquid sample from the original sample, and using the preset volume of liquid sample to dissolve a fixed amount of dry reagent mixture, which can ensure that the chemical components after dissolution have the same or similar concentration when performing different amplification reactions, such as the concentration difference is no more than 20%.

[0046] The specific implementation examples are all preferred embodiments of the present application and are not intended to limit the protection scope of the application. Therefore, any equivalent changes made based on the structure, shape, and principle of the present application should be included in the protection scope of the present application.

Claims

1. A liquid phase nucleic acid sample volume measuring consumable device, characterized in that: The invention comprises a sample collection tube (1), a metering component (2) connected to the sample collection tube (1), a sealing component (3) arranged between the sample collection tube (1) and the metering component (2), a reaction tube (6) connected to the metering component (2), and a cut-off component (7) arranged in the cavity of the consumable device, wherein the metering component (2) comprises a metering shell (20) connected to the sealing component (3), a metering tube (22) connected to the cut-off component (7) through an upper opening, and an opening and closing component (23) for sealing the lower end of the metering tube (22). The sample collection tube (1) is provided with an external connection structure on the outer side of one open end, the sample collection tube (1) is connected to the sealing component (3) via the external connection structure, the outer periphery of the metering shell (20) is provided with a fixing component (5) for fixing the reaction tube (6), the sample collection tube (1) is coupled to the heater (4), and the liquid sample in the sample tube is heated to generate a relatively high air pressure, under this high air pressure, the lower liquid is pushed into the metering tube (22) through the filter pad (32), the cut-off plate (70) and the guide tube (71).

2. A liquid phase nucleic acid sample volume measurement consumable device according to claim 1, characterized in that: The sealing assembly (3) comprises a sealing shell (30) and sealing gaskets (31) arranged at the upper and lower ends of the sealing shell (30); an internal connection structure is arranged inside the end of the sealing shell (30) close to the sample collection tube (1); the external connection structure of the sample collection tube (1) is connected to the internal connection structure of the sealing shell (30); and the external connection structure and the internal connection structure are threads or buckles.

3. A liquid phase nucleic acid sample volume measurement consumable device according to claim 2, characterized in that: The sealing component (3) is provided with a filter pad (32) therein. The filter pad (32) has the capability of filtering insoluble particles or the capability of adsorbing soluble non-nucleic acid inorganic molecules or organic molecules. The filter pad (32) is in the form of a membrane or a pad, and its matrix is ​​a mesh material or a porous granular material.

4. The liquid phase nucleic acid sample volume measuring consumable device according to claim 1, characterized in that: The truncation assembly (7) comprises a support plate (73), a truncation plate (70) located below the support plate (73), a flow guide tube (71) used to connect the truncation plate (70) and extend below the truncation plate (70) into the metering tube (22), and a truncation block (72) arranged below the truncation plate (70). The truncation plate (70) and the flow guide tube (71) are made of elastic material, and a through hole for the sample to flow is opened on the support plate (73).

5. A liquid phase nucleic acid sample volume measuring consumable device according to claim 4, characterized in that: The support plate (73) is arranged in the cavity of the metering component (2), or the support plate (73) is arranged in the cavity of the sealing component (3).

6. A liquid phase nucleic acid sample volume measuring consumable device according to claim 4, characterized in that: The truncated block (72) is a water-swellable polymer, and the water-swellable polymer includes a hydrogel.

7. A liquid phase nucleic acid sample volume measuring consumable device according to claim 1, characterized in that: The fixing assembly (5) comprises a fixing buckle (50) fixedly arranged on the outer periphery of the metering housing (20) and a clamping piece (51) fixedly arranged on the periphery of the upper end of the reaction tube (6).

8. The liquid phase nucleic acid sample volume measuring consumable device according to claim 1, characterized in that: The opening and closing assembly (23) comprises a twistable cover (230) arranged on one end of the metering tube (22) close to the reaction tube (6) and a stopper (231) sleeved on the twistable cover (230), the reaction tube (6) and the stopper (231) having no sliding fit, and the metering tube (22) and the metering housing (20) having no sliding fit.

9. The liquid phase nucleic acid sample volume measuring consumable device according to claim 1, characterized in that: A current limiting device (8) is provided above the cutoff component, and the current limiting device (8) is a heat-sensitive structure or a pressure-sensitive structure.

10. A method for measuring the volume of a liquid-phase nucleic acid sample, applied to a liquid-phase nucleic acid sample volume measuring consumable device according to any one of claims 1 to 9, characterized in that: The following steps are involved: S1, connecting a sample collection tube (1) containing a nucleic acid sample liquid to be detected to a sealing component (3) to form a fully sealed nucleic acid sample volume measurement consumable device; S2, coupling the heater (4) to the sample collection tube (1) and starting the heater (4), whereby the heater (4) can inactivate cells or viruses and release nucleic acid molecules; S3, the liquid sample in the sample tube is heated to generate a relatively high air pressure, under which the lower liquid is pushed into the metering tube (22) through the filter pad (32), the cut-off plate (70) and the flow guide tube (71); S4, when the liquid sample overflows in the metering tube (22), the cut-off block (72) absorbs the overflowing liquid sample and expands, thereby pushing the cut-off plate (70) and closing the through hole and / or the flow guide tube (71) of the support plate (73), thereby cutting off the passage of the liquid from the sample tube to the metering tube (22); S5, rotating the reaction tube (6) so that the reaction tube (6) drives the stopper (231) to rotate, twisting off the twistable cover (230), and opening the passage between the metering tube (22) and the reaction tube (6); S6. Under the action of gravity or centrifugal force, the preset volume of liquid sample in the metering tube (22) is transferred to the reaction tube (6).

Images