Integrated consumable device and method for nucleic acid sample preparation and volume measurement

Abstract

The present application relates to the field of nucleic acid detection technology, and in particular to an integrated consumable device and method for integrating nucleic acid sample preparation and volume measurement. First, a sample processing tube containing a liquid sample is docked to a mounting ring, and then a heater is used to heat the liquid sample in the sample processing tube, and a sufficiently high temperature is generated in the sample processing tube to lyse eukaryotic cells or prokaryotic cells or viruses in the sample, and release nucleic acids. The temperature can also be high enough to denature proteins and inactivate pathogens. The elevated temperature can generate sufficient internal gas pressure, which is sufficient to pass the liquid sample through the connecting groove and the three-way valve into the metering container. The liquid sample can fill the space of the metering container, and the liquid will contact the actuator to activate the actuator, causing the three-way valve to shift to close the liquid outflow of the sample processing tube, and at the same time open the channel between the metering container and the reaction tube, so that the metered sample is moved into the reaction tube connected to the docking ring.

Description

Technical Field

[0001] The present application relates to the field of nucleic acid detection technology, and in particular to an integrated consumable device and method for integrating nucleic acid sample preparation and volume measurement. Background Art

[0002] Molecular diagnostic and detection technologies based on nucleic acids are playing an increasingly important role in many fields. Nucleic acids are used in the detection of pathogenic microorganisms, species identification, species origin, diversity assessment, as well as applications and research work such as kinship, systematic evolution, and genetic engineering. These tasks all involve nucleic acid sample preparation. In the prior art, nucleic acid sample preparation involves multiple steps. Such as 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. In this process, people need to use centrifuges, pipettes, vortexers, etc. These operations need to be completed in the laboratory, and their operations are labor-intensive, cumbersome, and prone to errors. Summary of the invention

[0003] In view of the shortcomings of the prior art, the purpose of this application is to integrate a consumable device and method for nucleic acid sample preparation and volume measurement, which is used to solve the cumbersome operation problems of nucleic acid sample preparation and sample volume measurement before the detection reaction in the prior art.

[0004] The above-mentioned purpose of the present application is achieved through the following technical solutions: an integrated consumable device integrating nucleic acid sample preparation and volume measurement, comprising a flat sheet-shaped metering component, a sample processing tube placed vertically above the metering component, and a reaction tube for nucleic acid amplification reaction placed vertically below the metering component, the metering component comprising a metering container for measuring liquid volume and a fluid control board providing a channel for liquid flow, the fluid control board comprising a switching device for controlling liquid flow, the switching device comprising an actuator and a three-way valve coupled to the actuator, wherein after the metering container is filled with liquid, the flow of the liquid sample from the sample processing tube is terminated by activating the switching device, and the three-way valve can be designed as a cylindrical shape and allows any two of the three inlets / outlets to be connected.

[0005] Furthermore, the fluid control panel is provided with an L-shaped passage, a T-shaped placement slot and a C-shaped connecting slot. One end of the L-shaped passage is connected to the sample processing tube, and the other end is connected to the T-shaped placement slot. The end of the T-shaped placement slot away from the L-shaped passage is connected to the C-shaped connecting slot. Both ends of the C-shaped connecting slot are connected to the T-shaped placement slot. The actuator and the three-way valve are installed in the T-shaped placement slot and connected to the C-shaped connecting slot. The C-shaped connecting slot is connected to the metering container.

[0006] By adopting the above technical scheme, when using, first place the sample processing tube with the nucleic acid sample to be detected above the metering component, place the reaction tube below the fluid control board, and then use an ultrasonic generator or a heater to heat the liquid sample in the sample processing tube. The ultrasonic generator can be used for cell disruption and nucleic acid release. The heater can be an electromagnetic induction heater, a resistance heater (built-in or external), a semiconductor heater such as a Peltier heater, an infrared heater, a microwave heater, etc. A sufficiently high temperature will be generated in the sample processing tube to crack eukaryotic cells or prokaryotic cells or viruses, etc., and release nucleic acids. The temperature can also be high enough to cause protein denaturation and pathogen inactivation. In addition, the elevated temperature can generate enough pressure inside the sample processing tube, which is enough to pass the liquid sample through the L-shaped passage and the three-way valve into the metering container, and the metering container is a structure with a fixed volume, and the liquid sample can fill the space of the metering container. When the metering container (unit) is full, the liquid will contact the actuator to activate the actuator, and the actuator will push the three-way valve to shift, that is, the channel conversion, to close the liquid outflow from the sample processing tube, and at the same time open the channel between the metering container and the reaction tube, so that the measured sample can be moved into the reaction tube. After metering, the liquid sample of known volume is mixed with the required reagents in the reaction tube, which can maintain the consistency of the concentration of the reaction components in the reaction system between the reaction tubes, thereby ensuring the repeatability of the amplification reaction in the reaction tube. In this application, it is only necessary to heat the sample in the sample processing tube, drive the liquid flow by pressure, and drive the three-way valve to shift by the actuator, which greatly reduces the number of operating steps. The use of this device can achieve the effect of "sample in-result out", which is extremely practical.

[0007] Furthermore, the metering container includes a metering tube and an extrusion structure; the extrusion structure includes a relaxed compression spring fixedly arranged at one end of the metering tube away from the three-way valve, and an extrusion plate fixedly arranged at one end of the relaxed compression spring close to the three-way valve. The extrusion plate material may include metal, glass, rubber, plastic, Teflon, silicone, etc. When the pressure at both ends of the extrusion plate is unbalanced, it can move along the metering tube.

[0008] By adopting the above technical solution, when the metering container is full, the liquid will contact the actuator to activate the actuator and shift the three-way valve. At this time, the pressure on the metering container is reduced, and the compression spring is relaxed to release the liquid sample in the metering container, and a predetermined amount of liquid sample enters the reaction tube.

[0009] Furthermore, the metering component also includes a mounting ring fixedly arranged above the fluid control board and provided for docking the sample processing tube, a vertical groove is opened in the middle of the mounting ring, and the vertical groove is connected to the L-shaped passage. The metering component also includes a docking ring fixedly arranged below the fluid control board for installing the reaction tube, a through groove is opened in the middle of the docking ring and is connected to the T-shaped placement groove.

[0010] By adopting the above technical solution, the sample processing tube containing the nucleic acid sample to be detected is first connected to the installation ring, then inverted, and then the liquid sample in the sample processing tube is heated. The installation ring can prevent the sample from leaking under high pressure. The sample processing tube can also have openings at both ends. When one end is connected to the installation ring, the opening at the other end can be used for adding samples. After adding samples, the sample processing tube will be sealed to prevent the sample from leaking under high pressure. After the actuator pushes the three-way valve to shift, that is, the channel is converted, the sample will flow into the reaction tube in the docking ring along the C-type connection groove.

[0011] Furthermore, a filter pad is installed at the bottom of the mounting ring, and a sealing ring is arranged on the outer side of the filter pad, so that the filter pad is fixedly connected to the mounting ring.

[0012] By adopting the above technical solution, the filter pad is used to filter particles or sediments present in the heated sample, and the filter pad can be made of any material used for biological filtration. There are many filtering methods, such as based on size, charge, absorption, affinity binding and combinations of the above. The matrix material of the filter pad includes cellulose, nylon, resin, glass, metal, metal oxide or a combination thereof, which can be composed of a mesh, porous granules. The surface of these matrices can be functionally modified, such as carboxyl, amino, sulfonic, hydroxyl, benzyl, fatty, etc.

[0013] Furthermore, the actuator is made of a water-swellable polymer, and the water-swellable polymer includes a hydrogel.

[0014] By adopting the above technical solution, the material that can swell when encountering liquid can push the three-way valve to complete the transposition, that is, channel conversion. The water-absorbing and swellable polymer in this application includes hydrogel, which can be polyacrylamide or polyethylene oxide or carboxymethyl cellulose or modified starch or alginate or pectin. All water-soluble or hydrophilic polymers can form hydrogels 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, alginic 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 such as polyacrylic acid, polymethacrylic acid, polyacrylamide, poly-N-polyacrylamide, etc.

[0015] Furthermore, the metering container is communicated with the C-shaped connecting groove, an execution groove is provided above the fluid control plate, a hydrophobic membrane is installed in the execution groove, and the execution groove is communicated with the C-shaped connecting groove.

[0016] By adopting the above technical solution, the hydrophobic membrane is used to discharge the air in the fluid channel, thereby releasing the pressure; and at the same time prevent the aqueous solution from passing through.

[0017] Furthermore, it also includes a flow limiting device, which includes a thermosensitive structure or a pressure-sensitive structure. A pressure-sensitive structure, such as a pressure-sensitive membrane, can be placed between the hydrophobic membrane and the actuator; its function is that when the pressure difference on both sides of the pressure-sensitive membrane reaches a preset pressure, the device can conduct. The preset pressure can be above 1.5 atmospheres, or above 2.0 atmospheres, or above 2.5 atmospheres, or above 3 atmospheres. When the liquid sample flows through the metering tube through the C-type connecting groove and guides to the actuator, the air in the pipeline can be discharged through the hydrophobic membrane. When the liquid reaches the hydrophobic membrane and the pressure-sensitive structure, the liquid pressure is less than the preset pressure at this time, and the movement of the front end of the liquid will stop, but the metering tube connected to the C-type connecting groove will be filled with the liquid sample. When the liquid fills the metering tube, the pressure in the C-type connecting groove and the metering tube will increase until the pressure-sensitive structure is turned on, the actuator absorbs water and expands, and the three-way valve is pushed to shift or move up and down to achieve channel conversion. At this time, there is a pressure difference between the liquid in the metering tube and the space in the reaction tube, and the liquid in the metering tube flows to the reaction tube. The above operations achieve the purpose of automatic sampling, sample volume measurement, and pipetting.

[0018] Furthermore, the filter pad is in the form of a membrane or a pad, and its matrix is ​​a mesh material or a porous granular material.

[0019] By adopting the above technical solution, the porous structure of the filter pad can effectively intercept insoluble particles in the sample, such as cell debris, sediment, etc. If these particles enter the subsequent detection steps, they may interfere with the results of nucleic acid detection or even damage the detection instrument. Through the filtering effect of the filter pad, it can ensure that the liquid sample entering the metering container is purer, improving the accuracy and reliability of the detection.

[0020] Furthermore, a sample volume measurement method is provided, which uses any one of the above technical solutions to form an integrated consumable device for nucleic acid sample preparation and volume measurement, comprising the following steps:

[0021] S1, connecting the sample processing tube containing the liquid sample to the metering component;

[0022] S2, coupling a heater to the sample processing tube and heating the liquid sample in the sample processing tube through the heater, wherein the heater may be a resistance heater, an infrared heater, an electromagnetic heater, or a microwave heater;

[0023] S3, after the liquid sample in the sample processing tube is heated, a relatively high air pressure is generated in the tube cavity, and under this high air pressure, the liquid is pushed into the measuring container through the filter pad;

[0024] S4, the liquid sample fills the measuring container;

[0025] S5. Drive the three-way valve to allow the liquid sample in the metering container to flow into the reaction tube.

[0026] By adopting the above technical solution, the rapid preparation of samples is achieved by connecting the sample processing tube and the metering component. This method not only reduces the complexity and error rate of manual operation, but also improves the efficiency of the entire detection process. Using a variety of heating methods such as resistance heaters, infrared heaters, electromagnetic heaters or microwave heaters, the most suitable heating method can be selected according to the sample characteristics and detection requirements. Accurate heating control helps to ensure that there is an appropriate air pressure state in the sample processing tube, thereby effectively pushing the liquid sample through the filter pad into the metering container. This process avoids direct contact between the sample and the external environment, reduces the risk of cross contamination, and ensures the accuracy of the test results. The design of the metering container ensures that the liquid sample can be filled and reach the preset volume, thereby achieving accurate quantification of the sample. This is crucial for the subsequent nucleic acid detection steps, because the accurate sample volume directly affects the reliability and repeatability of the test results. By driving the three-way valve, the liquid sample in the metering container is transferred to the reaction tube. The automation of this step not only reduces the time and cost of manual operation, but also improves the accuracy and repeatability of the operation. The entire process, from sample processing to sample transfer to reaction system preparation, has achieved a high degree of automation and integration, which not only improves detection efficiency, but also reduces the impact of human factors on test results, making the nucleic acid detection process more standardized and normalized.

[0027] In order to prevent the sample from flowing into the metering tube too early, the integrated consumable device for nucleic acid sample preparation and volume metering of the present application is provided with a flow limiting device, which includes a thermosensitive structure or a pressure-sensitive structure. For example, a flow limiting device, such as a thermosensitive structure or a pressure-sensitive structure, is provided in the channel between the metering component and the sealing component or between the actuator and the hydrophobic membrane. The structure may be a plastic film that can be broken at a given temperature or a given air pressure. For example, 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, liquid can be allowed to flow through; or when the air pressure reaches 1.5 atmospheres or above, or 2.0 atmospheres or above, or 2.5 atmospheres or above, or 3 atmospheres or above, liquid can be allowed to flow through.

[0028] The reaction tube can be used with an instrument based on known technology to perform nucleic acid amplification reactions. Such instruments can achieve temperature control, light signal acquisition and data analysis.

[0029] In summary, the present application includes at least one of the following beneficial technical effects:

[0030] 1. The device integrates multiple steps such as sample processing, metering, and pipetting, greatly simplifying the operation process, reducing human intervention, and improving detection efficiency and accuracy. The liquid flow is automatically driven by heating, and the hydrogel actuator that swells when exposed to water controls the position of the three-way valve, realizing automatic metering and pipetting of samples, reducing the number of operation steps and dependence on peripheral auxiliary equipment.

[0031] 2. The metering container has a fixed volume, which can ensure the accurate amount of sample transferred each time, avoiding the errors and waste caused by improper human operation in traditional methods. At the same time, the design of the extrusion structure such as the relaxation compression spring and the extrusion plate ensures that the liquid sample in the metering container can be automatically filled or released quantitatively when the pressure changes.

[0032] 3. There is no need to use complex microfluidic structures, and there is no intermediate pipetting step. After the sample enters the sample processing tube, it does not leave the reaction consumable device until the nucleic acid amplification is completed. Compared with related technologies in this industry (such as reagent cartridges), except for the lyophilized reagents pre-placed in the reaction tube, the present invention does not pre-store liquid in the entire device; and the reaction liquid can be 100% from the sample itself. In terms of performance and design, production, and use costs, the product of the present invention has extremely high practicality. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is a schematic diagram of the overall structure in the embodiment;

[0034] Figure 2 is a cross-sectional view of the interior of the fluid control panel in the embodiment;

[0035] Figure 3 is a schematic diagram of the structure after the actuator moves in the embodiment;

[0036] Figure 4 is a schematic structural diagram of Embodiment 2;

[0037] Figure 5 is a schematic structural diagram of Embodiment 3;

[0038] Figure 6 It is a structural schematic diagram of Example 4.

[0039] Figure numerals: 1. fluid control plate; 21. mounting ring; 22. execution groove; 23. hydrophobic membrane; 25. sealing ring; 26. filter pad; 31. docking ring; 32. groove; 4. sample processing tube; 5. reaction tube; 51. flange; 6. metering container; 61. relaxed compression spring; 62. extrusion plate; 7. actuator; 71. three-way valve; 8. vertical groove; 82. L-type passage; 83. T-type placement groove; 84. C-type connecting groove; 9. flexible bubble; 91. cylinder plug. DETAILED DESCRIPTION

[0040] The present application is further described in detail below with reference to the accompanying drawings.

[0041] Example 1, reference Figure 1 , Figure 2 , an integrated consumable device integrating nucleic acid sample preparation and volume measurement, comprising a flat sheet-shaped metering component, a sample processing tube 4 vertically placed above the metering component, and a reaction tube 5 for nucleic acid amplification reaction vertically placed below the metering component, the metering component comprising a metering container 6 for measuring the volume of a liquid and a fluid control board 1 providing a channel for the flow of the liquid, the fluid control board 1 comprising a switch device for controlling the flow of the liquid, the switch device comprising an actuator 7 and a three-way valve 71 coupled to the actuator 7, wherein the metering container 6 automatically stops the outflow of the liquid sample from the sample processing tube 4 after being filled with liquid, an L-shaped passage 82, a T-shaped placement groove 83 and a C-shaped connection groove 84 are provided in the fluid control board 1, one end of the L-shaped passage 82 is connected to the T-shaped placement groove 83, the end of the T-shaped placement groove 83 away from the L-shaped passage 82 is connected to the C-shaped connection groove 84, both ends of the C-shaped connection groove 84 are connected to the T-shaped placement groove 83, the actuator 7 and the three-way valve 71 are installed in the T-shaped placement groove 83, and the C-shaped connection groove 84 is connected to the metering container 6.

[0042] When using, refer to Figure 3 First, place the sample processing tube 4 above the metering component, place the reaction tube 5 below the fluid control board 1, and then use a heater to heat the liquid sample in the sample processing tube 4 on the metering component (the heater can be an electromagnetic induction heater, a resistance heater (built-in or external), a Peltier heater, an infrared heater or a microwave heater, etc.), and generate a sufficiently high temperature in the sample processing tube 4 to lyse eukaryotic cells or prokaryotic cells or viruses, etc., and release nucleic acids. The temperature can also be high enough to denature proteins and inactivate pathogens. In addition, the increased temperature can cause a pressure higher than the external environment to be generated inside the sample processing tube 4, which is sufficient to push the liquid sample through the L-shaped passage 82 and through the three-way valve 71 into the metering container 6. The metering container 6 is a structure with a fixed volume, and the liquid sample can fill the space of the metering container 6. When the metering container 6 (unit) is full, the liquid will contact the actuator 7 to activate the actuator 7, which will push the three-way valve 71 to shift to close the outflow of the liquid from the sample processing tube 4 and open the channel between the metering container 6 and the reaction tube 5 at the same time. At this time, the pressure applied to the liquid by the spring in the metering container is greater than the pressure applied to the liquid by the gas in the reaction tube 5, so that the measured sample is moved into the reaction tube 5, dissolving the reagent dry ball placed in the reaction tube 5, so that the reaction tube 5 can be directly used for nucleic acid amplification reaction. In this embodiment, it is only necessary to heat the sample in the sample processing tube 4 to automatically complete multiple operations such as nucleic acid release, sample filtration, volume measurement, and pipetting, which greatly reduces the manual operation steps and has extremely high practicality.

[0043] In this embodiment, the metering container 6 includes a metering tube and an extrusion structure. The extrusion structure includes a relaxed compression spring 61 fixedly arranged on one end of the metering tube away from the three-way valve 71, and an extrusion plate 62 fixedly arranged on the end of the relaxed compression spring 61 close to the three-way valve 71.

[0044] When the metering container 6 is full, the liquid will contact the actuator 7 and activate the actuator 7 to shift the three-way valve 71. At this time, since the pressure of the metering container 6 is greater than the pressure in the reaction tube, the compression spring 61 is relaxed to release the liquid sample in the metering container 6, and a predetermined amount of liquid will enter the reaction tube 5.

[0045] In this embodiment, the metering assembly includes a mounting ring 21 fixedly disposed above the fluid control board 1 and provided for docking the sample processing tube 4 . A vertical groove 8 is provided in the middle of the mounting ring 21 , and the vertical groove 8 is connected to an L-shaped passage 82 .

[0046] First, the sample processing tube 4 containing the nucleic acid sample to be detected is connected to the mounting ring 21, then inverted, and then the liquid sample in the sample processing tube 4 is heated. The setting of the mounting ring 21 can make the sample processing tube 4 seal and fit to the metering component to prevent the sample from flowing out under high pressure. The sealing method can be a thread plus a sealing ring. The sample processing tube 4 can also have openings at both ends. When one end is connected to the mounting ring 21, the opening at the other end can be used for sample addition. After sample addition, the sample processing tube 4 will be sealed to prevent the sample from leaking under high pressure.

[0047] In this embodiment, a filter pad 26 is installed at the bottom of the installation ring 21 , a sealing ring 25 is arranged on the outer side of the filter pad 26 , and the filter pad 26 is fixedly connected to the installation ring 21 .

[0048] The filter pad 26 is used to filter particles or sediments present after the sample is heated. The filter pad 26 can be made of any material used for biological filtration. There are many ways of filtering; for example, based on size, charge, absorption, affinity binding, and combinations of the above.

[0049] In this embodiment, the actuator 7 is made of a water-swellable polymer, which includes a hydrogel, or other water-swellable polymers such as polyacrylamide, polyethylene oxide, carboxymethyl cellulose, modified starch, alginate, or pectin.

[0050] The material that expands when encountering liquid can push the three-way valve 71 to complete the channel conversion.

[0051] In the present application, a flange 51 is fixedly provided on the upper edge of the reaction tube 5 , and a matching groove 32 is provided on the inner side of the docking ring 31 to prevent the reaction tube 5 from falling off when being inserted into the docking ring 31 .

[0052] In this embodiment, the metering assembly further includes a docking ring 31 fixedly disposed below the fluid control plate 1 , which is helpful for installing the reaction tube 5 , and a through groove is opened in the middle thereof and is connected to the T-shaped placement groove 83 .

[0053] After the actuator 7 pushes the three-way valve 71 , the sample will flow into the reaction tube 5 in the docking ring 31 along the C-shaped connecting groove.

[0054] An execution groove 22 is provided above the fluid control plate 1 , a hydrophobic membrane 23 is installed in the execution groove 22 , and the execution groove 22 is communicated with the C-shaped connection groove 84 .

[0055] The hydrophobic membrane 23 is used to discharge the air in the fluid channel, thereby releasing the pressure; at the same time it prevents the aqueous solution from passing through.

[0056] The filter pad 26 is a porous structure and has the capability of filtering insoluble particles or adsorbing dissolved inorganic salt ions or non-nucleic acid organic molecules.

[0057] The porous structure of the filter pad 26 can effectively intercept insoluble particles in the sample, such as cell debris, sediment, etc. If these particles enter the subsequent detection steps, they may interfere with the results of nucleic acid detection. Through the filtering effect of the filter pad 26, it can be ensured that the liquid sample entering the metering container 6 is purer, and the accuracy and reliability of the detection are improved.

[0058] A sample volume measurement method is applied to any one of the above technical solutions to an integrated consumable device integrating nucleic acid sample preparation and volume measurement, comprising the following steps:

[0059] S1, connecting the sample processing tube 4 containing the liquid sample to the metering component;

[0060] S2, coupling an ultrasonic generator or a heater to the sample processing tube 4 and heating the liquid sample in the sample processing tube 4 through the heater, which may be a resistance heater, an infrared heater, an electromagnetic heater or a microwave heater;

[0061] S3, after the liquid sample in the sample processing tube is heated, a relatively high air pressure is generated in the tube cavity, and under this high air pressure, the liquid is pushed into the metering container 6 through the filter pad 26;

[0062] S4, the liquid sample fills the measuring container 6;

[0063] S5 , driving the three-way valve 71 , and moving the liquid sample in the measuring container 6 into the reaction tube 5 .

[0064] By connecting the sample processing tube 4 and the metering component, not only the complexity and error rate of manual operation are reduced, but also the efficiency of the entire detection process is improved. By using a variety of heating methods such as resistance heaters, infrared heaters, electromagnetic heaters or microwave heaters, the most suitable heating method can be selected according to the sample characteristics and detection requirements. The heating power can be pre-calibrated and automatically adjusted by electronic circuits. Accurate heating control helps to ensure that the sample quickly reaches a high pressure state at an appropriate temperature and prevents the air pressure from being too high, thereby effectively pushing the liquid sample through the filter pad 26 into the metering container 6. In order to prevent the sample from flowing into the metering tube too early, a flow limiting device is provided in the channel between the actuator and the hydrophobic membrane. The flow limiting device is a heat-sensitive or pressure-sensitive structure, which can be a plastic film that can be broken at a given temperature or a given air pressure. With such a structure, 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, liquid can be allowed to flow through; or when the air pressure reaches 1.5 atmospheres or above, or 2.0 atmospheres or above, or 2.5 atmospheres or above, or 3 atmospheres or above, liquid can be allowed to flow through.

[0065] Driven by high air pressure, the liquid sample is pushed into the metering container 6 through the filter pad 26. This process avoids direct contact between the sample and the external environment, reduces the risk of cross contamination, and ensures the accuracy of the test results. The design of the metering container 6 ensures that the liquid sample can be filled and reach the preset volume, thereby achieving accurate quantification of the sample volume. This is crucial for the subsequent nucleic acid detection steps, because the accurate sample size directly affects the reliability and repeatability of the test results. By driving the three-way valve 71, the liquid sample in the metering container 6 is transferred to the reaction tube 5. The automation of this step not only reduces the time and cost of manual operation, but also improves the accuracy and repeatability of the operation. The entire process from sample processing to sample transfer to the reaction tube 5 is highly automated and integrated, which not only improves the detection efficiency, but also reduces the impact of human factors on the test results, making the nucleic acid detection process more standardized and normalized.

[0066] Example 2, reference Figure 4 The difference between this embodiment and embodiment 1 is that the metering component is arranged above the fluid control board 1, and is connected to the C-type connection groove 84 through the fluid control board 1. By arranging the metering component above the fluid control board 1, the situation in the metering component can be more clearly understood during use. Embodiment 2 is similar to embodiment 1, but the metering component is arranged on the fluid control board 1, and is connected to the C-type connection groove 84 through the fluid control board 1. The workflow of this embodiment is similar to that of embodiment 1, but the metering process is more intuitive, and the user can directly observe the metering situation from the top, which improves the visibility of the metering and the convenience of operation.

[0067] Example 3, reference Figure 5 , the difference between this embodiment and embodiment 1 and embodiment 2 is that the metering container 6 uses an enlarged C-type connecting groove 84 for metering. The extrusion structure is changed to a flexible bubble 9 fixedly connected to the fluid control board 1 at the end of the hydrophobic membrane 23. The working process is: the sample processing tube 4 is connected to the heater, the liquid enters the enlarged C-type connecting groove 84, and the gas is discharged from the hydrophobic membrane 23. When the liquid contacts the hydrophobic membrane 23, the liquid cannot pass through the hydrophobic membrane 23. At this time, the hydrogel actuator 7 that expands when exposed to water is activated, and the three-way valve 71 immediately closes the channel of the sample processing tube 4, and the flow stops. The liquid in the enlarged C-type connecting groove 84 reaches a predetermined amount. By squeezing the flexible bubble 9, the gas is reversely entered into the enlarged C-type connecting groove and the liquid sample is pushed into the reaction tube. Therefore, the cooperation of the fixed volume catheter and the hydrophobic membrane 23 provides another metering method.

[0068] Example 4, reference Figure 6 The difference between this embodiment and Embodiment 1, Embodiment 2 and Embodiment 3 is that the actuator is changed into an extrusion rod 7, and the extrusion structure is replaced with a cylinder plug 91. When the cylinder plug 91 pushes the sample, it further ensures that the liquid sample in the metering container 6 can be accurately and residue-free released into the reaction tube 5.

[0069] Specific implementation process: A mounting ring 21 is provided above the sample processing fluid control board 1 for plugging in the sample processing tube 4, and lysing cells or viruses through a heating mechanism to release nucleic acids. During the heating process, the internal pressure pushes the liquid sample through the connecting groove into the three-way valve 71. The three-way valve 71 is a key component, and its cylindrical design allows for the connection of any two of the three inlets / outlets. When the metering container 6 is filled with liquid samples, the actuator 7 is activated, and the three-way valve 71 immediately closes the channel of the sample processing tube 4, and at the same time opens the channel between the metering container 6 and the reaction tube 5, and opens the extrusion structure such as the cylinder plug 91 to achieve accurate metering and pipetting of samples.

[0070] In addition, the device is also equipped with a filter pad 26 and a sealing ring 25 to filter impurities in the sample and prevent liquid leakage.

[0071] During use, users only need a few simple steps to complete sample lysis, filtration, metering, pipetting and preparation before amplification reaction, without the need for complicated operations and peripheral equipment assistance. This device not only improves detection efficiency, but also reduces operation difficulty and cost.

[0072] The amplification reaction can be a constant temperature reaction or a variable temperature reaction. The reaction tube 5 cooperates with the temperature and optical device to achieve the purpose of nucleic acid amplification. The reaction tube 5 can be used in conjunction with an instrument based on known technology to perform nucleic acid amplification reactions. Such instruments can achieve temperature control, optical signal acquisition and data analysis.

[0073] In Example 4, the actuator 7 is changed into an extrusion cylinder, and the spring is replaced by a cylinder plug 91. The extrusion cylinder is controlled by air pressure or hydraulic pressure to achieve precise indexing of the three-way valve 71. The working process is similar to that of the aforementioned Example 3, but after the metering container 6 is filled, the extrusion cylinder receives a signal and starts working. The cylinder plug 91 pushes to close the channel of the sample processing tube 4 and open the channel between the metering container 6 and the reaction tube 5. The liquid in the metering container 6 is released into the reaction tube 5, which improves the control accuracy and response speed of the actuator 7 and increases the stability of the device.

[0074] In the above embodiment, a pressure-sensitive structure, such as a pressure-sensitive membrane, may be disposed between the hydrophobic membrane 23 and the actuator 7; its function is that when the pressure difference on both sides of the pressure-sensitive structure reaches a preset pressure, the device can conduct. The preset pressure may be above 1.5 atmospheres, or above 2.0 atmospheres, or above 2.5 atmospheres, or above 3 atmospheres. When the liquid sample flows through the metering tube through the C-type connecting groove 84 and is directed to the actuator 7, the air in the pipeline may be discharged through the hydrophobic membrane 23. When the liquid reaches the hydrophobic membrane 23 and the pressure-sensitive structure, the liquid pressure is less than the preset pressure, and the front end of the liquid stops moving, but the metering tube connected to the C-type connecting groove 84 will be filled with the liquid sample. When the liquid fills the metering tube, the pressure in the tube will increase until the pressure-sensitive structure is turned on, the actuator 7 absorbs water and expands, and the three-way valve 71 is pushed to shift or move up and down to realize channel conversion. At this time, the liquid in the metering tube can flow to the reaction tube, achieving the purpose of automatic sampling, sample volume measurement, and pipetting.

[0075] The sample volume measurement of the present application refers to obtaining a preset volume of liquid sample from the original sample. The use of the preset volume of liquid sample is to ensure that the chemical components have the same or similar concentrations when performing different amplification reactions after dissolving a fixed amount of dry reagent mixture, such as a concentration difference of no more than 20%.

[0076] The embodiments of the specific implementation methods of this application are all preferred embodiments of this application, and are not intended to limit the scope of protection of the application. Therefore, all equivalent changes made based on the structure, shape, and principle of this application should be included in the scope of protection of this application.

Claims

1. An integrated consumable device for nucleic acid sample preparation and volume measurement, characterized in that: The invention comprises a flat sheet-shaped metering component, a sample processing tube (4) vertically placed above the metering component, and a reaction tube (5) for performing a nucleic acid amplification reaction vertically placed below the metering component. The metering component comprises a metering container (6) for measuring the volume of a liquid and a fluid control board (1) for providing a channel for the flow of the liquid. The fluid control board (1) comprises a switch device for controlling the flow of the liquid. The switch device comprises an actuator (7) and a three-way valve (71) coupled to the actuator (7). After the metering container is filled with liquid, the switch device is activated to stop the liquid sample from flowing out of the sample processing tube (4). The fluid control panel (1) is provided with an L-shaped passage (82), a T-shaped placement groove (83) and a C-shaped connection groove (84); one end of the L-shaped passage (82) is connected to the sample processing tube (4), and the other end is connected to the T-shaped placement groove (83); the end of the T-shaped placement groove (83) away from the L-shaped passage (82) is connected to the C-shaped connection groove (84); both ends of the C-shaped connection groove (84) are connected to the T-shaped placement groove (83); the actuator (7) and the three-way valve (71) are installed in the T-shaped placement groove (83) and connected to the C-shaped connection groove (84); and the C-shaped connection groove (84) is connected to the metering container (6).

2. The integrated consumable device for nucleic acid sample preparation and volume measurement according to claim 1, characterized in that: The metering container (6) comprises a metering tube and an extrusion structure.

3. The integrated consumable device for nucleic acid sample preparation and volume measurement according to claim 1, characterized in that: The metering assembly further comprises a mounting ring (21) arranged above the fluid control plate (1) for docking the sample processing tube (4), a vertical groove (8) being provided in the middle of the mounting ring (21), the vertical groove (8) being communicated with the L-shaped passage (82), and a docking ring (31) being arranged below the fluid control plate (1) for mounting the reaction tube (5), a through groove being provided in the middle of the docking ring (31) being communicated with the lower end of the T-shaped placement groove (83).

4. The integrated consumable device for nucleic acid sample preparation and volume measurement according to claim 3, characterized in that: A filter pad (26) is installed at the bottom of the installation ring (21), a sealing ring (25) is arranged on the outside of the filter pad (26), and the filter pad (26) is fixedly connected to the installation ring (21).

5. The integrated consumable device for nucleic acid sample preparation and volume measurement according to claim 1, characterized in that: The actuator (7) is made of a water-swellable polymer, and the water-swellable polymer includes a hydrogel.

6. The integrated consumable device for nucleic acid sample preparation and volume measurement according to claim 1, characterized in that: It also includes a current limiting device, which includes a heat-sensitive structure or a pressure-sensitive structure.

7. The integrated consumable device for nucleic acid sample preparation and volume measurement according to claim 1, characterized in that: The metering container (6) is in communication with the C-shaped connecting groove (84); an execution groove (22) is provided above the fluid control plate (1); a hydrophobic membrane (23) is installed in the execution groove (22); and the execution groove (22) and the C-shaped connecting groove (84) are in communication.

8. The integrated consumable device for nucleic acid sample preparation and volume measurement according to claim 4, characterized in that: The filter pad (26) is in the form of a membrane or a pad, and its matrix is ​​a mesh material or a porous granular material.

9. A sample volume measurement method, applied to the integrated consumable device for nucleic acid sample preparation and volume measurement according to any one of claims 1 to 8, characterized in that: The following steps are involved: S1, connecting the sample processing tube (4) containing the liquid sample to the metering component; S2, coupling a heater to the sample processing tube (4) and heating the liquid sample in the sample processing tube (4) by the heater, wherein the heater includes a semiconductor heater, a resistance heater, an infrared heater, an electromagnetic heater, or a microwave heater; S3, after the liquid sample in the sample processing tube is heated, a relatively high air pressure is generated in the tube cavity, and under this high air pressure, the liquid is pushed into the metering container (6) through the filter pad (26); S4, the liquid sample fills the measuring container (6); S5, driving the three-way valve (71) of the switch device to stop the liquid sample from flowing out of the sample processing tube (4), and at the same time moving the liquid sample in the measuring container (6) into the reaction tube.

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