One-step multi-layer serial nucleic acid extraction device

Abstract

The invention relates to the field of nucleic acid extraction, and discloses a one-step multi-layer tandem nucleic acid extraction device, which comprises a sealing cover, a buffer solution tandem tube structure for separating nucleic acid and a swab, the buffer solution tandem tube structure for separating nucleic acid comprises a lysis solution tube, a binding solution tube, a washing solution tube I, a washing solution tube II, an eluent tube I and an eluent tube II, one end of the lysate pipe is connected with a combined liquid pipe, the other end of the combined liquid pipe is connected with a washing liquid pipe I, the other end of the washing liquid pipe I is connected with a washing liquid pipe II, the other end of the washing liquid pipe II is connected with an eluent pipe I or an eluent pipe II, and the eluent pipe I and the eluent pipe II can be replaced. Due to the integrated design of the swab arranged in the buffer solution series tube structure for separating nucleic acid, a swab sample sequentially passes through the buffer solution series tube and the sealing film, the whole process treatment of nucleic acid extraction is completed in the device, and all the components are tightly matched through the puncturable sealing film structure, so that the independent sealing of a reagent and the smoothness of swab insertion are ensured.

Description

Technical Field

[0001] This invention relates to the field of nucleic acid extraction, specifically to a one-step multilayer tandem nucleic acid extraction device. Background Technology

[0002] Nucleic acids, as a key component of life, play a crucial role in diagnostics in the biomedical field. Their detection process typically involves two core steps: nucleic acid extraction and nucleic acid amplification. Nucleic acid extraction aims to isolate and purify DNA / RNA molecules from complex biological samples, laying the foundation for subsequent accurate nucleic acid amplification and detection. Currently, mainstream nucleic acid extraction technologies include magnetic bead method and centrifuge column method.

[0003] From an operational perspective, many one-step multilayer tandem nucleic acid extraction devices have cumbersome operating procedures. For example, the operation of reagents provided by traditional kits often requires a lot of time, and cross-contamination can easily occur during the operation due to operating methods or experimental environment issues. This places stringent requirements on the experimental skills of medical personnel and the testing environment, which not only increases the difficulty and burden of the operators, but may also affect the quality of nucleic acid extraction and the accuracy of subsequent test results. To address this, we propose a one-step multilayer tandem nucleic acid extraction device. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a one-step multilayer tandem nucleic acid extraction device, which solves the aforementioned problems.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution: a one-step multilayer tandem nucleic acid extraction device, comprising a sealing cap, a buffer series tube structure for separating nucleic acids, and a swab, wherein one end of the buffer series tube structure for separating nucleic acids is connected to the sealing cap, the swab is snapped into the buffer series tube structure for separating nucleic acids, and the tail of the swab is connected to the sealing cap; The buffer series tube structure for separating nucleic acids includes a lysis buffer tube, a binding buffer tube, a washing buffer tube 1, a washing buffer tube 2, an elution buffer tube 1, and an elution buffer tube 2. One end of the lysis buffer tube is connected to the binding buffer tube, the other end of the binding buffer tube is connected to the washing buffer tube 1, and the other end of the washing buffer tube 1 is connected to the washing buffer tube 2. The end of the washing liquid tube 2 that is away from the washing liquid tube 1 is connected to either the elution liquid tube 1 or the elution liquid tube 2. Both the elution liquid tube 1 and the elution liquid tube 2 correspond to the connection end of the washing liquid tube 2. The elution liquid tube 1 and the elution liquid tube 2 can be replaced. The end of the pyrolysis fluid tube opposite to the binding fluid tube is connected to the sealing cap.

[0006] Preferably, an internally threaded tube is connected through the sealing cap, one end of which is fixedly connected to the inner plane of the sealing cap, and a connecting tube is fixedly connected to the plane of the sealing cap opposite to the internally threaded tube, the connecting tube communicating with the internally threaded tube.

[0007] Preferably, a puncture seal is inserted into one end of the internally threaded tube that connects to the connecting tube, and the tail of the swab is inserted into the connecting tube and fits against the puncture seal.

[0008] Preferably, the sealing cap is fixedly connected to an externally threaded pipe on the side opposite to the internally threaded pipe, and the externally threaded pipe is on the outside of the connecting pipe.

[0009] Preferably, one end of the lysis fluid tube is fixedly connected to an internally threaded connecting tube, and a puncture seal is inserted into the end of the internally threaded connecting tube connected to the lysis fluid tube. The externally threaded tube is inserted into the internally threaded connecting tube and threadedly connected to it. The puncture seal is located between the end of the externally threaded tube away from the sealing cap and the internally threaded connecting tube. The end of the swab away from the sealing cap passes through the internally threaded connecting tube, the puncture seal, the lysis fluid tube, and the externally threaded connecting tube in sequence.

[0010] Preferably, the end of the lysis fluid tube opposite to the internally threaded connecting tube is fixedly connected to an externally threaded connecting tube, and the end of the binding fluid tube is fixedly connected to an internally threaded connecting tube. A puncture seal is inserted into the end of the internally threaded connecting tube connected to the binding fluid tube. The externally threaded connecting tube is inserted into the internally threaded connecting tube and threadedly connected to it. The puncture seal is located between the end of the externally threaded connecting tube opposite to the lysis fluid tube and the internally threaded connecting tube. The end of the swab opposite to the externally threaded connecting tube passes through the internally threaded connecting tube, the puncture seal, and the binding fluid tube in sequence.

[0011] Preferably, one end of the binding fluid tube opposite to the internally threaded connecting tube two is fixedly connected to an externally threaded connecting tube two, one end of the washing fluid tube one is fixedly connected to an internally threaded connecting tube three, and a puncture seal two is inserted into the end of the internally threaded connecting tube three connected to the washing fluid tube one. The externally threaded connecting tube two is inserted into the internally threaded connecting tube three and threadedly connected to the internally threaded connecting tube three. The puncture seal two is located between the end of the externally threaded connecting tube two opposite to the binding fluid tube and the internally threaded connecting tube three. The end of the swab opposite to the externally threaded connecting tube two passes through the internally threaded connecting tube three, the puncture seal two, and the washing fluid tube one in sequence.

[0012] Preferably, one end of the washing liquid tube 1, away from the internally threaded connecting tube 3, is fixedly connected to an externally threaded connecting tube 3; one end of the washing liquid tube 2 is fixedly connected to an internally threaded connecting tube 6; a puncture seal 2 is inserted into the end of the internally threaded connecting tube 6 that connects to the washing liquid tube 2; the externally threaded connecting tube 3 is inserted into the internally threaded connecting tube 6 and threadedly connected to it; the puncture seal 2 is located between the end of the externally threaded connecting tube 3 away from the washing liquid tube 1 and the internally threaded connecting tube 6; and the end of the swab away from the externally threaded connecting tube 3 passes through the internally threaded connecting tube 6, the puncture seal 2, and the washing liquid tube 2 in sequence.

[0013] Preferably, the end of the washing liquid tube 2 opposite to the internally threaded connecting tube 6 is fixedly connected to the externally threaded connecting tube 4, and the end of the elution liquid tube 1 is fixedly connected to the internally threaded connecting tube 4. A puncture seal 2 is inserted into the end of the internally threaded connecting tube 4 that is connected to the elution liquid tube 1. The externally threaded connecting tube 4 is inserted into the internally threaded connecting tube 4 and threadedly connected to the internally threaded connecting tube 4. The puncture seal 2 is located between the end of the externally threaded connecting tube 4 opposite to the washing liquid tube 2 and the internally threaded connecting tube 4. The end of the swab opposite to the washing liquid tube 2 is inside the elution liquid tube 1.

[0014] Preferably, one end of the eluent tube two is fixedly connected to an internally threaded connecting tube five, which corresponds to the externally threaded connecting tube four.

[0015] Compared with the prior art, the advantages of the present invention are as follows: A one-step multilayer tandem nucleic acid extraction device is provided, which has the following advantages: This invention, through its integrated structural design, enables the entire process of sample processing, from collection to nucleic acid elution, to be completed within the same device. This eliminates the need for complex equipment and tube transfer operations, effectively simplifying the operation steps, shortening the extraction time, and reducing the risk of cross-contamination caused by tube transfer. It also improves the efficiency and accuracy of nucleic acid extraction. Furthermore, the components are tightly fitted through a puncture-resistant sealing structure and threaded connections, ensuring independent sealing of reagents and smooth flow of liquid. This invention is suitable for nucleic acid extraction needs of various sample types. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the structure of the present invention; Figure 2 This is a cross-sectional schematic diagram of the buffer series tube structure for separating nucleic acids according to the present invention; Figure 3 for Figure 2 A magnified view of part A in the diagram; Figure 4 for Figure 2 A magnified view of part B in the diagram.

[0017] In the diagram: 1. Sealing cap; 2. Internally threaded tube; 3. Externally threaded tube; 4. Connecting tube; 5. Swab; 6. Puncture seal 1; 7. Lysis fluid tube; 8. Binding fluid tube; 9. Washing fluid tube 1; 10. Eluent tube 1; 11. Eluent tube 2; 12. Internally threaded connecting tube 1; 13. Externally threaded connecting tube 1; 14. Internally threaded connecting tube 2; 15. Externally threaded connecting tube 2; 16. Internally threaded connecting tube 3; 17. Externally threaded connecting tube 3; 18. Internally threaded connecting tube 4; 19. Internally threaded connecting tube 5; 20. Puncture seal 2; 21. Washing fluid tube 2; 22. Internally threaded connecting tube 6; 23. Externally threaded connecting tube 4. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] Please see Figure 1-4 The present invention provides a technical solution: a one-step multilayer tandem nucleic acid extraction device, comprising a sealing cap 1, a buffer series tube structure for separating nucleic acids and a swab 5, wherein one end of the buffer series tube structure for separating nucleic acids is connected to the sealing cap 1, the swab 5 is snapped into the buffer series tube structure for separating nucleic acids, and the tail of the swab 5 is connected to the sealing cap 1. The buffer series tube structure for separating nucleic acids includes a lysis buffer tube 7, a binding buffer tube 8, a washing buffer tube 1 9, a washing buffer tube 2 21, an elution buffer tube 1 10, and an elution buffer tube 2 11. One end of the lysis buffer tube 7 is connected to the binding buffer tube 8, the other end of the binding buffer tube 8 is connected to the washing buffer tube 1 9, and the other end of the washing buffer tube 1 9 is connected to the washing buffer tube 2 21. The end of the washing liquid tube 21 that is away from the washing liquid tube 9 is connected to the elution liquid tube 10 or the elution liquid tube 21. The elution liquid tube 10 and the elution liquid tube 21 are both connected to the end of the washing liquid tube 21. The elution liquid tube 10 and the elution liquid tube 21 can be replaced. The end of the pyrolysis fluid tube 7 that is away from the binding fluid tube 8 is connected to the sealing cap 1.

[0020] Furthermore, an internally threaded tube 2 is connected through the sealing cap 1. One end of the internally threaded tube 2 is fixedly connected to the inner plane of the sealing cap 1. A connecting tube 4 is fixedly connected to the plane of the sealing cap 1 opposite to the internally threaded tube 2. The connecting tube 4 is connected to the internally threaded tube 2. The internally threaded tube 2 is used to connect the sealing cap 1 and the threaded extraction tube or syringe. The connecting tube 4 is used to connect the swab 5.

[0021] Furthermore, a puncture seal 6 is inserted into one end of the internally threaded tube 2 that connects to the connecting tube 4. The tail of the swab 5 is inserted into the connecting tube 4 and fits against the puncture seal 6. When not punctured, the puncture seal 6 blocks the internally threaded tube 2 and the connecting tube 4. After puncture, the puncture seal 6 forms a through-hole structure with the hollow swab 5. The sampling head of the swab 5 is modified and optimized by a special process to specifically adsorb nucleic acids and can interact with different buffer solutions in the series tube through the puncture seal 6. The diameter of the swab 5 is the same as the diameter of the puncture seal 6. When the swab 5 passes through the series tube, the puncture seal... The seal 6 can tightly seal the swab 5 and the series tube to prevent liquid leakage. In addition, during the insertion or mixing and shaking process of the swab 5, the buffer solution that has already interacted with the swab 5 is prevented from entering the next series tube and mixing with the next buffer solution. The sampling head of the swab 5 is modified and optimized by a special process to specifically adsorb nucleic acids. The special process modification and optimization includes, but is not limited to, functional fillers (silica powder, hydroxy / carboxyl magnetic beads, silicone membrane) and ultra-high molecular weight polyethylene sintering molding process, poly(3,4-dihydroxy-L-phenylalanine) (L-DOPA) and nylon / flocked swab modification. The puncture seal 6 includes, but is not limited to, one or more combinations of silicone, rubber, polypropylene, Teflon, aluminum foil membrane or other microporous filter membranes or PE sieve plates.

[0022] Furthermore, an externally threaded pipe 3 is fixedly connected to the side of the sealing cap 1 that is opposite to the internally threaded pipe 2. The externally threaded pipe 3 is located outside the connecting pipe 4 and is used to connect the sealing cap 1 and the pyrolysis liquid pipe 7.

[0023] Furthermore, one end of the lysis fluid tube 7 is fixedly connected to an internally threaded connecting tube 12. A puncture seal 20 is inserted into the end of the internally threaded connecting tube 12 that is connected to the lysis fluid tube 7. An externally threaded tube 3 is inserted into the internally threaded connecting tube 12 and threadedly connected to it. The puncture seal 20 is located between the end of the externally threaded tube 3 away from the sealing cap 1 and the internally threaded connecting tube 12. The end of the swab 5 away from the sealing cap 1 passes through the internally threaded connecting tube 12, the puncture seal 20, the lysis fluid tube 7, and the externally threaded connecting tube 13 in sequence. The externally threaded tube 3 and the internally threaded connecting tube 12 are used to connect the sealing cap 1 and the lysis fluid tube 7. The puncture seal 20 in the internally threaded connecting tube 12 is used to block the lysis fluid tube 7 and the externally threaded tube 3 and connect the swab 5.

[0024] Furthermore, an externally threaded connecting pipe 13 is fixedly connected to one end of the pyrolysis fluid pipe 7 away from the internally threaded connecting pipe 12, and an internally threaded connecting pipe 24 is fixedly connected to one end of the binding fluid pipe 8. A puncture seal 20 is inserted into the end of the internally threaded connecting pipe 24 that connects to the binding fluid pipe 8. The externally threaded connecting pipe 13 is inserted into the internally threaded connecting pipe 24 and threadedly connected to it. The puncture seal 20 is located at the end of the externally threaded connecting pipe 13 away from the pyrolysis fluid pipe 7 and the internally threaded connecting pipe 12. Between the two connecting pipes 14, the end of the swab 5 away from the external threaded connecting pipe 13 passes through the internal threaded connecting pipe 14, the puncture seal 20, and the binding fluid pipe 8 in sequence. The external threaded connecting pipe 13 and the internal threaded connecting pipe 14 are used to connect the lysis fluid pipe 7 and the binding fluid pipe 8. The puncture seal 20 in the internal threaded connecting pipe 14 is used to block the lysis fluid pipe 7 and the binding fluid pipe 8 and connect the swab 5. The lysis fluid pipe 7 and the binding fluid pipe 8 can be the same series pipe, that is, the lysis and binding reactions are carried out simultaneously in the same pipe.

[0025] Furthermore, an external threaded connecting tube 15 is fixedly connected to one end of the combined fluid tube 8 away from the internal threaded connecting tube 14, and an internal threaded connecting tube 16 is fixedly connected to one end of the washing fluid tube 9. A puncture seal 20 is inserted into the end of the internal threaded connecting tube 16 that connects to the washing fluid tube 9. The external threaded connecting tube 15 is inserted into the internal threaded connecting tube 16 and threadedly connected to it. The puncture seal 20 is located between the end of the external threaded connecting tube 15 away from the combined fluid tube 8 and the internal threaded connecting tube 16. The end of the swab 5 away from the external threaded connecting tube 15 passes through the internal threaded connecting tube 16, the puncture seal 20, and the washing fluid tube 9 in sequence. The external threaded connecting tube 15 and the internal threaded connecting tube 16 are used to connect the combined fluid tube 8 and the washing fluid tube 9. The puncture seal 20 in the internal threaded connecting tube 16 is used to block the combined fluid tube 8 and the washing fluid tube 9 and connect the swab 5.

[0026] Furthermore, an externally threaded connecting pipe 3 17 is fixedly connected to one end of the washing liquid tube 1 9 away from the internally threaded connecting pipe 3 16. An internally threaded connecting pipe 6 22 is fixedly connected to one end of the washing liquid tube 2 21. A puncture seal 2 20 is inserted into the end of the internally threaded connecting pipe 6 22 that connects to the washing liquid tube 2 21. The externally threaded connecting pipe 3 17 is inserted into the internally threaded connecting pipe 6 22 and threadedly connected to it. The puncture seal 2 20 is located between the end of the externally threaded connecting pipe 3 17 away from the washing liquid tube 1 9 and the internally threaded connecting pipe 6 22. The end of the swab 5 away from the externally threaded connecting pipe 3 17 passes through the internally threaded connecting pipe 6 22 in sequence. The threaded connecting tube 22, the puncture seal 20, and the washing liquid tube 21 are used to connect the washing liquid tube 19 and the washing liquid tube 21. The puncture seal 20 in the internal threaded connecting tube 22 is used to block the washing liquid tube 21 and the washing liquid tube 19 and connect the swab 5. The washing liquid punctures the puncture seal 16 in the internal threaded tube 2 and enters the head of the swab 5 through the tail of the swab 5. The number of washing liquid series tubes of the swab 5, the washing liquid tube 19, and the washing liquid tube 21 can be reduced or increased according to the specific project requirements. The total number of washing liquid series tubes is ≥1.

[0027] Furthermore, one end of the washing liquid tube 21 opposite to the internally threaded connecting tube 6 22 is fixedly connected to an externally threaded connecting tube 4 23. One end of the eluent tube 10 is fixedly connected to an internally threaded connecting tube 4 18. A puncture seal 20 is inserted into the end of the internally threaded connecting tube 4 18 that connects to the eluent tube 10. The externally threaded connecting tube 4 23 is inserted into the internally threaded connecting tube 4 18 and threadedly connected to it. The puncture seal 2 20 is located on the externally threaded connecting tube 4 23 opposite to the internally threaded connecting tube 4 18. Between one end of the washing liquid tube 21 and the internal threaded connecting tube 4 18, the end of the swab 5 away from the washing liquid tube 21 is inside the elution tube 10. The external threaded connecting tube 4 23 and the internal threaded connecting tube 4 18 are used to connect the washing liquid tube 21 and the elution tube 10. The puncture seal 20 inside the internal threaded connecting tube 4 18 is used to block the washing liquid tube 21 and the elution tube 10 and connect the swab 5. The elution tube 10 is filled with elution liquid and can be removed.

[0028] Furthermore, one end of the elution tube 2 11 is fixedly connected to an internally threaded connecting tube 5 19, which corresponds to the externally threaded connecting tube 4 23. The internally threaded connecting tube 5 19 is used to connect the elution tube 2 11 and the washing tube 2 21. The elution tube 2 11 contains elution solution, which can elute the cleaned nucleic acid adsorbed on the swab 5 head into the elution solution. The elution tube 2 11 can be removed separately.

[0029] Structural Description: Sealing cap 1: The whole is a threaded cap with a puncture-proof structure. The upper end has an internal thread or Luer interface, and the lower end has an external thread and a insertion hole. It is used to connect the threaded extraction tube or syringe, fix the tail of swab 5, and is the top of the sealing device. It is connected to the buffer series tube structure for separating nucleic acids and is the core connector at the top. Internal thread tube 2: Tubular structure, one end is fixed to the inner plane of the sealing cap 1, with internal threads, used to connect the sealing cap 1 and the threaded extraction tube or syringe, providing a channel for liquid injection; External threaded tube 3: A tubular structure located outside the connecting tube 4, with external threads on the outer wall, used for threaded connection with the internal threaded connecting tube 12 of the pyrolysis fluid tube 7 to fix the sealing cap 1 to the pyrolysis fluid tube 7. Connecting tube 4: A tubular structure that connects to the internally threaded tube 2. It is used to insert the swab 5 into the tail, fix the swab, and serve as a channel for liquid flow. Swab 5: It consists of a head and a hollow rod. The head is made of flocked, nylon or other materials. The rod is hollow and ends at the top of the head. It is used to collect samples (such as oral, nasal, and pharyngeal swabs). Liquid can enter from the tail and flow out from the head to realize sample processing and liquid circulation. Puncture seal 6: A punctureable sealing structure that is inserted into the connection end of the internally threaded tube 2 and the connecting tube 4. When not punctured, it prevents the two tubes from being broken apart. After puncturing, it forms a through-hole structure with the hollow swab 5, ensuring liquid flow while maintaining a seal. Lysis buffer tube 7: A tubular container filled with lysis buffer, with internal threaded connecting tube 12 and external threaded connecting tube 13 connected at both ends to contain the sample and release the nucleic acid from the sample; Binding fluid tube 8: A tubular container filled with binding fluid, with internally threaded connecting tube 2 14 and externally threaded connecting tube 2 15 connected at both ends, used to allow the released nucleic acid to specifically bind with the components of the binding fluid; Washing liquid tube 19: A tubular container with internal threaded connecting tube 316 and external threaded connecting tube 317 connected at both ends, used to hold washing liquid and work with washing liquid tube 21 to clean impurities from the head of swab 5. Elution tube 10: A tubular container filled with elution solution, with one end connected to internally threaded connecting tube 4 18, used to elute the nucleic acid adsorbed on the head of swab 5. It can be removed separately to collect the elution solution. Elution tube 21: A tubular container filled with elution solution. One end is connected to internally threaded connection tube 519, which is used to elute the nucleic acid adsorbed on the head of swab 5. It can be removed separately to collect the elution solution and is interchangeable with elution tube 10. Internal threaded connecting tube 12: tubular structure, one end is fixed to the lysate tube 7, with internal threads, and a puncture seal 20 is inserted inside for connection with the external threaded tube 3, so as to realize the connection between the sealing cap 1 and the lysate tube 7 and isolate the two. External threaded connecting pipe 13: a tubular structure, fixed to the end of the pyrolysis fluid pipe 7 away from the internal threaded connecting pipe 12, with external threads on the outer wall, used to connect with the internal threaded connecting pipe 14 to fix the pyrolysis fluid pipe 7 and the binding fluid pipe 8. Internal threaded connecting tube 14: tubular structure, one end is fixed to the binding fluid tube 8, with internal threads, and a puncture seal 20 inserted inside, used to connect with external threaded connecting tube 13, so as to connect the pyrolysis fluid tube 7 and the binding fluid tube 8 and isolate the two. External threaded connecting pipe 2 15: tubular structure, fixed to the end of the connecting liquid pipe 8 away from the internal threaded connecting pipe 2 14, with external threads on the outer wall, used to connect with the internal threaded connecting pipe 3 16 to fix the connecting liquid pipe 8 and the washing liquid pipe 1 9. Internal threaded connecting tube 316: tubular structure, one end is fixed to washing liquid tube 19, with internal thread, and a puncture seal 20 inserted inside, used to connect with external threaded connecting tube 215, to realize the connection between the combined liquid tube 8 and the washing liquid tube 19 and isolate the two. Internal threaded connecting tube 4 18: tubular structure, one end is fixed to the eluent tube 1 10, with internal threads, and a puncture seal 20 is inserted inside, used to connect with external threaded connecting tube 4 23, so as to connect the washing liquid tube 2 21 and the eluent tube 1 10 and isolate the two. Internal threaded connecting tube 519: tubular structure, fixed to one end of eluent tube 21, with internal thread for corresponding connection with external threaded connecting tube 423, realizing the connection between eluent tube 21 and washing liquid tube 21. Puncture seal 20: A puncture-proof sealing structure that is inserted into each internally threaded connecting tube and located between the connecting ends of two adjacent tubes. When not punctured, it blocks the adjacent tubes. After puncturing, it adheres to the swab 5 to maintain a seal, ensuring that the reagents in each tube are independent and that the liquid can flow through the swab. Washing liquid tube 21: A tubular container with internal threaded connecting tube 6 22 and external threaded connecting tube 4 23 connected at both ends, used to hold washing liquid. It works with washing liquid tube 1 9 to clean impurities from the head of swab 5. The quantity can be increased or decreased as needed. Together with washing liquid tube 1 9, it forms a series washing liquid tube. Internal threaded connecting tube 6 22: tubular structure, one end is fixed to washing liquid tube 2 21, with internal thread, and puncture seal 2 20 is inserted inside, used to connect with external threaded connecting tube 3 17, so as to connect washing liquid tube 1 9 and washing liquid tube 2 21 and isolate the two. External threaded connecting pipe 4 23: a tubular structure, fixed to the end of washing liquid pipe 21 opposite to the internal threaded connecting pipe 6 22, with external threads on the outer wall, used to connect with internal threaded connecting pipe 4 18 or internal threaded connecting pipe 5 19, to fix washing liquid pipe 21 to elution liquid pipe 10 or elution liquid pipe 2 11.

[0030] Working principle: During the sample collection and initial assembly stage, the sampling personnel collect the samples to be tested by hand using a swab according to standard procedures, such as oropharyngeal swabs and nasopharyngeal swabs commonly used in clinical diagnosis. After collection, the tail of the swab 5 is precisely inserted into the connecting tube 4 of the sealing cap 1, ensuring that the swab and the connecting tube are tightly fitted and that it is in contact with the puncture seal 6 in the internally threaded tube 2 to prevent sample leakage and external contamination. At the same time, the sealing cap 1 is threadedly connected to the internally threaded connecting tube 12 of the lysis fluid tube 7 through the externally threaded tube 3. During operation, the thread tightening specifications must be followed to ensure the sealing and stability of the connection. At this time, the puncture seal 20 inside the internal threaded connecting pipe 12 is located between the external threaded pipe 3 and the pyrolysis fluid pipe 7. Then, the series pipes are assembled in sequence. The external threaded connecting pipe 13 of the pyrolysis fluid pipe 7 is threaded to the internal threaded connecting pipe 24 of the binding fluid pipe 8. The external threaded connecting pipe 25 of the binding fluid pipe 8 is threaded to the internal threaded connecting pipe 36 of the washing fluid pipe 19. The external threaded connecting pipe 317 of the washing fluid pipe 19 is threaded to the internal threaded connecting pipe 62 of the washing fluid pipe 21. When each adjacent pipe is connected, the puncture seal 20 inside the corresponding internal threaded connecting pipe plays an isolation role.Finally, select either eluent tube 10 or eluent tube 21 according to the requirements. If eluent tube 10 is selected, connect the external threaded connector 423 of eluent tube 21 to the internal threaded connector 418 of eluent tube 10. If eluent tube 21 is selected, connect it to the external threaded connector 423 through the internal threaded connector 519. Then proceed to the step-by-step processing stage. Apply external force to the sealing cap 1 or the tail of swab 5 to push swab 5 slowly downward. The head of swab 5 first penetrates the internal threaded connector 12, and then punctures the puncture seal 20, successfully entering the lysis fluid tube 7. At this time, the puncture seal 20 is tightly attached. The outer wall of swab 5 remains sealed. The lysis buffer in lysis buffer tube 7 quickly comes into contact with the sample at the head of swab 5. If lysis buffer tube 7 and binding buffer tube 8 are connected in series, the lysis and binding reactions occur simultaneously. If they are independent tubes, after the lysis reaction is fully completed and the cell membranes and nuclear membranes of the sample cells are destroyed, and the nucleic acids are released into the solution, swab 5 is continuously pushed so that its head penetrates the puncture seal 20 in the threaded connecting tube 24 corresponding to binding buffer tube 8 and enters binding buffer tube 8. The specific binding components in the binding buffer bind to the nucleic acids, causing the nucleic acids to be adsorbed onto the head of swab 5. Then, the head of swab 5 is pushed down further, sequentially... The puncture seal 20 inside the internal threaded connecting tube 3 16 of the washing liquid tube 19 and the puncture seal 20 inside the internal threaded connecting tube 6 22 of the washing liquid tube 21 enters the washing liquid tube 21. At this time, connect the threaded extraction tube or syringe to the internal threaded tube 2 and inject the washing liquid. The washing liquid forms a through hole with the hollow swab 5 after being punctured by the puncture seal 1 6. The tail of the swab 5 flows into the head, rinsing impurities in the head. The insertion and insertion of the swab 5 prevents the washing liquid that has been in contact from entering other tubes. After washing is completed, if the connection is to the eluent tube 10, push the head of the swab 5 to puncture the internal threaded connecting tube 4 18. The puncture seal 20 is inserted into the elution tube 10. If it needs to be replaced with the elution tube 21, the elution tube 10 can be removed. The elution tube 21 is then connected to the external threaded connecting tube 4 23 via the internal threaded connecting tube 5 19. The swab head 5 is then pushed into the elution tube 21. The low ionic strength solution and other components in the elution solution disrupt the binding force between the nucleic acid and the swab, causing the nucleic acid to dissolve in the elution solution. Finally, either the elution tube 10 or the elution tube 21 is removed individually. The nucleic acid in the elution solution is collected by gravity drainage, centrifugation, syringe pressing, or pipette aspiration for subsequent PCR amplification and related detection.

[0031] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A one-step multi-layered serial nucleic acid extraction device, characterized in that, The application relates to a nucleic acid separation buffer series pipe structure and a swab, and belongs to the technical field of nucleic acid separation. The nucleic acid separation buffer series pipe structure comprises a lysis solution pipe (7), a binding solution pipe (8), a washing solution pipe I (9), a washing solution pipe II (21), an elution solution pipe I (10) and an elution solution pipe II (11). The elution solution pipe I (10) and the elution solution pipe II (11) are arranged in the washing solution pipe II (21) in a replaceable mode. The lysis solution pipe (7) is connected with the sealing cover (1) at one end.

2. The one-step multilayered serial nucleic acid extraction device according to claim 1, wherein, The sealing cover (1) is internally connected with a threaded pipe (2) in a penetrating mode, one end of the threaded pipe (2) is fixedly connected with the sealing cover (1) in a planar mode, and the sealing cover (1) is fixedly connected with a communication pipe (4) in a planar mode at a position away from the threaded pipe (2).

3. The one-step multi-layered serial nucleic acid extraction device of claim 2, wherein, The tail of the swab (5) is inserted into the communication pipe (4) and is attached to the puncture sealing member I (6).

4. The one-step multilayered serial nucleic acid extraction device of claim 2, wherein, The sealing cover (1) is fixedly connected with a threaded pipe (3) in a planar mode at a position away from the threaded pipe (2), and the threaded pipe (3) is arranged outside the communication pipe (4).

5. The one-step multi-layered serial nucleic acid extraction device of claim 4, wherein, One end of the lysis solution pipe (7) is fixedly connected with a threaded connection pipe I (12), the threaded connection pipe I (12) is internally inserted with a puncture sealing member II (20) at a position connected with the lysis solution pipe (7), the threaded pipe (3) is inserted into the threaded connection pipe I (12) and is threadedly connected with the threaded connection pipe I (12), the puncture sealing member II (20) is arranged between one end of the threaded pipe (3) away from the sealing cover (1) and the threaded connection pipe I (12), and one end of the swab (5) away from the sealing cover (1) is sequentially penetrated through the threaded connection pipe I (12), the puncture sealing member II (20), the lysis solution pipe (7) and a threaded connection pipe II (13).

6. The one-step multi-layered serial nucleic acid extraction device of claim 5, wherein, The lysis solution pipe (7) is fixedly connected with an outer threaded connection pipe one (13) at one end away from the inner threaded connection pipe one (12), one end of the binding solution pipe (8) is fixedly connected with an inner threaded connection pipe two (14), a puncture sealing piece two (20) is inserted in one end of the inner threaded connection pipe two (14) connected with the binding solution pipe (8), the outer threaded connection pipe one (13) is inserted in the inner threaded connection pipe two (14) and is threadedly connected with the inner threaded connection pipe two (14), the puncture sealing piece two (20) is between one end of the outer threaded connection pipe one (13) away from the lysis solution pipe (7) and the inner threaded connection pipe two (14), one end of the swab (5) away from the outer threaded connection pipe one (13) penetrates the inner threaded connection pipe two (14), the puncture sealing piece two (20) and the binding solution pipe (8) in sequence.

7. The one-step multi-layered serial nucleic acid extraction device according to claim 6, wherein, The binding solution pipe (8) is fixedly connected with an outer threaded connection pipe two (15) at one end away from the inner threaded connection pipe two (14), one end of the washing solution pipe one (9) is fixedly connected with an inner threaded connection pipe three (16), a puncture sealing piece two (20) is inserted in one end of the inner threaded connection pipe three (16) connected with the washing solution pipe one (9), the outer threaded connection pipe two (15) is inserted in the inner threaded connection pipe three (16) and is threadedly connected with the inner threaded connection pipe three (16), the puncture sealing piece two (20) is between one end of the outer threaded connection pipe two (15) away from the binding solution pipe (8) and the inner threaded connection pipe three (16), one end of the swab (5) away from the outer threaded connection pipe two (15) penetrates the inner threaded connection pipe three (16), the puncture sealing piece two (20) and the washing solution pipe one (9) in sequence.

8. The one-step multi-layered serial nucleic acid extraction device according to claim 7, wherein, The washing solution pipe one (9) is fixedly connected with an outer threaded connection pipe three (17) at one end away from the inner threaded connection pipe three (16), one end of the washing solution pipe two (21) is fixedly connected with an inner threaded connection pipe six (22), a puncture sealing piece two (20) is inserted in one end of the inner threaded connection pipe six (22) connected with the washing solution pipe two (21), the outer threaded connection pipe three (17) is inserted in the inner threaded connection pipe six (22) and is threadedly connected with the inner threaded connection pipe six (22), the puncture sealing piece two (20) is between one end of the outer threaded connection pipe three (17) away from the washing solution pipe one (9) and the inner threaded connection pipe six (22), one end of the swab (5) away from the outer threaded connection pipe three (17) penetrates the inner threaded connection pipe six (22), the puncture sealing piece two (20) and the washing solution pipe two (21) in sequence.

9. The one-step multi-layered serial nucleic acid extraction device of claim 8, wherein, The washing liquid pipe two (21) is fixedly connected with an outer threaded connection pipe four (23) at one end away from the inner threaded connection pipe six (22), one end of the elution liquid pipe one (10) is fixedly connected with an inner threaded connection pipe four (18), a puncture sealing piece two (20) is inserted in one end of the inner threaded connection pipe four (18) connected with the elution liquid pipe one (10), the outer threaded connection pipe four (23) is inserted in the inner threaded connection pipe four (18) and is in threaded connection with the inner threaded connection pipe four (18), the puncture sealing piece two (20) is between one end of the outer threaded connection pipe four (23) away from the washing liquid pipe two (21) and the inner threaded connection pipe four (18), one end of the swab (5) away from the washing liquid pipe two (21) is in the elution liquid pipe one (10).

10. The one-step multi-layered serial nucleic acid extraction device of claim 9, wherein, One end of the elution liquid pipe two (11) is fixedly connected with an inner threaded connection pipe five (19), the inner threaded connection pipe five (19) corresponds to the outer threaded connection pipe four (23).

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