A sampling tube and application method for nucleic acid amplification and detection without opening the lid

Abstract

The present invention provides a sampling tube and an application method for realizing nucleic acid amplification and detection without opening the lid. The sampling tube includes a tube body and a tube cap; on the upper side of the outer wall of the tube body, a first protruding ring and a second protruding ring are provided from top to bottom; on the lower part of the inner wall of the tube cap, a first concave ring and a second concave ring are provided from top to bottom; the first protruding ring is adapted to the first concave ring and the second concave ring for sealing the tube body and the tube cap; the second protruding ring is adapted to the second concave ring for sealing the tube body and the tube cap; inside the tube cap, a waste liquid pool, a partitioned liquid storage structure, and a needle inserting structure are respectively provided from top to bottom; the needle inserting structure is used for piercing the partitioned liquid storage structure; a liquid discharge channel is provided between the waste liquid pool and the tube body. When inverted, the liquid drains from the tube body to the waste liquid pool, and when upright, the liquid remains in the waste liquid pool. The present invention can, without opening the lid, realize the step-by-step release of the reaction reagents in the tube cap into the reaction system by pressing the tube cap step by step, realize nucleic acid amplification and detection without opening the lid, and avoid the risk of aerosol contamination between samples and contamination of the testing personnel.

Description

Technical Field

[0001] The present invention relates to the technical field of nucleic acid detection. Specifically, it particularly relates to a sampling tube and an application method for realizing nucleic acid amplification and detection without opening the lid. Background Art

[0002] With the rapid development of molecular biology technology, microbial identification technology has also developed rapidly. Nucleic acid detection based on polymerase chain reaction (PCR) is one of the important technical means for virus and bacteria detection.

[0003] For the PCR detection of virus and bacteria nucleic acids, it usually needs to be carried out by trained detection personnel in a professional PCR laboratory. The professional PCR laboratory sets up different rooms with pressure differences according to the experimental steps of the PCR reaction to separately carry out reagent preparation, sample pretreatment, and amplification analysis. Based on the professional design of the PCR laboratory and the protection of personnel, the infection risk of detection personnel and the risk of cross - contamination between samples can be reduced to a certain extent. However, since it is still necessary to repeatedly open the sampling tube and reagent tube to transfer samples and / or reagents after sampling, the risks of personnel infection and sample cross - contamination cannot be completely avoided. Summary of the Invention

[0004] In order to overcome the problems in the prior art, the purpose of the present invention is to provide a sampling tube and an application method for realizing nucleic acid amplification and detection without opening the lid.

[0005] A sampling tube for realizing nucleic acid amplification and detection without opening the lid includes a tube body and a tube cap; on the upper side of the outer wall of the tube body, a first protruding ring and a second protruding ring are provided from top to bottom; on the lower part of the inner wall of the tube cap, a first concave ring and a second concave ring are provided from top to bottom; the first protruding ring is adapted to the first concave ring and the second concave ring for sealing the tube body and the tube cap; the second protruding ring is adapted to the second concave ring for sealing the tube body and the tube cap; inside the tube cap, a waste liquid pool, a partitioned liquid storage structure, and a needle inserting structure are respectively provided from top to bottom; the needle inserting structure is used to pierce the partitioned liquid storage structure; a liquid discharge channel is provided between the waste liquid pool and the tube body. When inverted, the liquid drains from the tube body to the waste liquid pool, and when upright, the liquid remains in the waste liquid pool.

[0006] The partitioned liquid storage structure is provided with a partition ring and an outer ring, and is divided into an inner area and an outer area. A sealing film is provided at the bottom of the partitioned liquid storage structure;

[0007] The needle inserting structure includes a disc - shaped base and a needle. The needle is fixed on the disc - shaped base. When the tube cap approaches the tube body, the disc - shaped base is supported by the tube orifice of the tube body, and the needle is used to pierce the sealing film.

[0008] The lengths of the needles corresponding to the inner and outer areas are different, which is used to release the liquid in the partitioned liquid storage structure step by step.

[0009] The sampling tube body has a larger upper part and a smaller lower part, with a conical bottom that can be inserted into the reaction wells of a conventional PCR instrument. The diameter of the open end is 1 cm.

[0010] One end of the drainage channel on the tube cap is located below the liquid separation and storage structure, and the other end is located near the inner wall of the tube cap close to the cap top to prevent the reaction liquid from flowing back after entering the waste liquid chamber.

[0011] The cross-section of the raised parts of the first protruding ring and the second protruding ring is semi-circular, and the cross-section of the sunken parts of the first concave ring and the second concave ring is semi-circular. The four have the same radius.

[0012] The center distance between the first protruding ring and the second protruding ring is the same as the center distance between the first concave ring and the second concave ring.

[0013] The separation ring and the outer ring are concentrically distributed, and the outer side and the top surface of the outer ring are connected to the inner wall and the top surface of the tube cap.

[0014] The 4 long hollow needles and 1 short hollow needle of the needle insertion structure are distributed in a plum blossom shape on the disc-shaped base, and the 1 short hollow needle is located at the center of the circle. The disc-shaped base is provided with round holes corresponding to the needles to facilitate the reaction reagent to flow out after the needles penetrate the sealing film.

[0015] The length difference between the 4 long hollow needles and the short hollow needle is consistent with the center distance between the first protruding ring and the second protruding ring.

[0016] The edge of the disc-shaped base of the needle insertion structure is recessed inward, and the cross-section is semi-circular, which meshes with the sealing rubber ring.

[0017] An application method of the sampling tube described above. The reaction reagent A is contained inside the sampling tube body; the reaction reagent B is contained between the separation ring and the outer ring of the liquid separation and storage structure, and the reaction reagent C is contained inside the separation ring.

[0018] The steps are as follows: After the sample is added to the sampling tube, cover the tube cap. After the reaction, the magnetic beads in the reaction reagent A are adsorbed on the bottom of the tube body by the external magnet at the bottom of the tube body. Rotate the whole sampling tube 180° clockwise to make the waste liquid completely enter the waste liquid pool through the drainage channel. Then rotate the sampling tube 180° clockwise again, press the tube cap down until the first protrusion ring is completely engaged with the second concave ring, and the long hollow needle in the needle insertion structure punctures the sealing film to release the reaction reagent B into the tube body. Remove the external magnet at the bottom of the tube body; after the reaction, rotate the tube body 90° counterclockwise and apply an external magnetic force on the left side wall to adsorb the magnetic beads; keep applying the external magnetic force on the left side wall continuously and rotate the tube body 90° clockwise, press the tube cap down until the first protrusion ring is completely engaged with the first concave ring and the second protrusion ring is completely engaged with the second concave ring, and the short hollow needle in the needle insertion structure punctures the sealing film to release the reaction reagent C into the tube body for reaction.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: Through the technical solution of the present invention, after adding the sample, the reagents required for the reaction can be added step by step without opening the lid to complete the whole nucleic acid detection process. Brief Description of the Drawings

[0020] Figure 1 It is a schematic front view structural diagram of the initial state of a sampling tube for nucleic acid amplification and detection without opening the lid according to the present invention.

[0021] Figure 2 It is a schematic front view structural diagram of a sampling tube for nucleic acid amplification and detection without opening the lid according to the present invention when adding the reaction reagent B.

[0022] Figure 3 It is a schematic front view structural diagram of a sampling tube for nucleic acid amplification and detection without opening the lid according to the present invention when adding the reaction reagent C.

[0023] Figure 4 It is a schematic top view structural diagram of a puncture device of a sampling tube for nucleic acid amplification and detection without opening the lid according to the present invention and a partial enlarged view of the meshing part of the base and the sealing rubber ring.

[0024] Figure 5 It is a schematic bottom view structural diagram of a partitioned liquid storage structure in the tube cap of a sampling tube for nucleic acid amplification and detection without opening the lid according to the present invention.

[0025] Figure 6 It is a schematic top view structural diagram of the cross-section of the waste liquid chamber of the tube cap of a sampling tube for nucleic acid amplification and detection without opening the lid according to the present invention.

[0026] In the figure, there are a tube body 10, a first protruding ring 11, a second protruding ring 12, a tube cap 20, a first concave ring 21, a second concave ring 22, a septum 23, a prism 24, a partitioned liquid storage structure 30, a partition ring 31, an outer ring 32, a sealing film 33, a needle inserting structure 40, a disc-shaped base 41, a long hollow needle 42, a short hollow needle 43, and a sealing rubber ring 44. Detailed implementation mode

[0027] To facilitate the understanding of the present invention, the present invention will be described more comprehensively below in conjunction with relevant attached drawings and embodiments. The described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments.

[0028] As Figures 1 to 6 shown, the present invention provides a sampling tube for realizing nucleic acid amplification and detection without opening the lid, including a sampling tube body 10. The outer part of the top end of the sampling tube body is provided with a tube cap 20. Inside the tube cap 20, there is a partitioned liquid storage structure 30, and below the partitioned liquid storage structure 30 is a needle inserting structure 40.

[0029] On the outer surface of the sampling tube body 10 near the opening end, there are a first protruding ring 11 and a second protruding ring 12.

[0030] Below the inner wall of the tube cap 20, there are a first concave ring 21 and a second concave ring 22. There is a protruding prism 24 on the right outer wall. Inside the prism 24 is a liquid discharge channel, and the space between the top inner wall and the septum 23 is a waste liquid chamber.

[0031] The partitioned liquid storage structure 30 includes a partition ring 31 and an outer ring 32 connected to the septum 23 inside the tube cap 20, and a sealing film 33 at the lower end for the needle inserting structure 40 to puncture.

[0032] The needle inserting structure 40 includes a disc-shaped base 41, 4 long hollow needles 42, 1 short hollow needle 43, and a sealing rubber ring 44.

[0033] Furthermore, the sampling tube body 10 is larger at the top and smaller at the bottom, and the bottom is conical, which can be inserted into the reaction holes of a conventional PCR instrument. The diameter near the opening end is 1 cm.

[0034] Furthermore, the opening end of the liquid discharge channel on the tube cap 20 is located below the partitioned liquid storage structure 30, and the other end is located at the inner wall near the cap top of the tube cap 20 to prevent the reaction reagent from flowing back after entering the waste liquid chamber.

[0035] It should be noted that in this embodiment, when the tube cap 20 is tightened, the sampling tube is rotated clockwise by 180° to an inverted state, so that the reaction reagent A after processing the sample enters the waste liquid chamber through the liquid discharge channel. Then continue to rotate clockwise by 180° to the initial position to prevent the reaction reagent A from flowing back from the waste liquid chamber.

[0036] Further, the relative position control between the tube cap 20 and the sampling tube body 10 is achieved through the engagement of the first concave ring 21 and the second concave ring 22 of the tube cap 20 with the first protruding ring 11 and the second protruding ring 12. The cross-section of the protruding parts of the first protruding ring 11 and the second protruding ring 12 is semi-circular, and the cross-section of the recessed parts of the first concave ring 21 and the second concave ring 22 is semi-circular. The four have the same radius. The center distance between the first protruding ring 11 and the second protruding ring 12 is the same as the center distance between the first concave ring 21 and the second concave ring 22.

[0037] Further, the partition ring 31 and the outer ring 32 included in the partition liquid storage structure 30 are concentrically distributed, and the outer side and the middle partition of the outer ring 32 are connected to the inner wall and the middle partition of the tube cap 20.

[0038] Further, the four long hollow needles 42 and one short hollow needle 43 of the needle insertion structure 40 are distributed in a plum blossom shape on the disc-shaped base 41, and the one short hollow needle 43 is located at the center of the circle; circular holes are provided at the positions corresponding to the needles on the disc-shaped base 41 to facilitate the reaction reagent to flow out after the needles penetrate the sealing film 33; the length difference between the four long hollow needles 42 and the short hollow needle 43 is consistent with the center distance between the first protruding ring 11 and the second protruding ring 12. The edge of the disc-shaped base 41 of the needle insertion structure 40 is recessed inward, and the cross-section is semi-circular, which engages with the sealing rubber ring 44.

[0039] It should be noted that as Figure 2 , when the tube cap 20 is pressed until the first protruding ring 11 and the second concave ring 22 are engaged with each other, the long hollow needle 42 in the needle insertion structure 40 will pierce the sealing film 33 and release the reaction reagent B into the sampling tube; when the tube cap 20 is further pressed until the first protruding ring 11 and the first concave ring 21, and the second protruding ring 12 and the second concave ring 22 are engaged respectively, the short hollow needle 43 in the needle insertion structure 40 will pierce the sealing film 33 and release the reaction reagent C into the sampling tube; thus, the step-by-step release of the reaction reagent B and the reaction reagent C is realized.

[0040] Further, the reaction reagent A is contained inside the sampling tube body 10.

[0041] Further, the reaction reagent B is contained between the partition ring 31 and the outer ring 32 of the partition liquid storage structure 30, and the reaction reagent C is contained inside the partition ring 31.

[0042] It should be noted that the reaction reagents A / B / C can be filled with appropriate reagents according to needs and reaction steps.

[0043] Application Example

[0044] A sampling tube that enables nucleic acid amplification and detection without opening the lid, and its specific usage steps are as follows: After the sample is added to the sampling tube, cover the tube lid. After reacting for a certain period of time, the magnetic beads in reagent A are adsorbed at the bottom of the tube body by the external magnetic force at the bottom of the tube body. Rotate the entire sampling tube 180° clockwise so that the waste liquid completely enters the waste liquid pool through the drainage channel. Then rotate the sampling tube 180° clockwise again, press the tube lid down until the first protruding ring is completely engaged with the second concave ring, and the long hollow needle in the needle insertion structure punctures the sealing film to release reagent B into the tube body, and remove the external magnet at the bottom of the tube body. After reacting for a certain period of time, rotate the tube body 90° counterclockwise and apply an external magnetic force on the left side wall to adsorb the magnetic beads; keep applying the external magnetic force on the left side wall continuously and rotate the tube body 90° clockwise, press the tube lid down until the first protruding ring is completely engaged with the first concave ring and the second protruding ring is completely engaged with the second concave ring, and the short hollow needle in the needle insertion structure punctures the sealing film to release reagent C into the tube body. After reacting for a certain period of time, observe the reaction result through other equipment.

[0045] The above content is a further detailed description of the present invention in combination with specific preferred implementation manners, and it cannot be determined that the specific implementation of the present invention is only limited to these descriptions. For those skilled in the art, without departing from the concept of the present invention, several simple deductions or substitutions can still be made, which all belong to the scope protected by the present invention.

Claims

1. A sampling tube that enables nucleic acid amplification and detection without opening the lid, characterized in that, It includes a tube body (10) and a tube cap (20); On the upper side of the outer wall of the tube body (10), a first protruding ring (11) and a second protruding ring (12) are provided from top to bottom; On the lower part of the inner wall of the tube cap, a first concave ring (21) and a second concave ring (22) are provided from top to bottom; The first protruding ring (11) is adapted to the first concave ring (21) and the second concave ring (22) for sealing the tube body (10) and the tube cap (20); The second protruding ring (12) is adapted to the second concave ring (22) for sealing the tube body (10) and the tube cap (20); Inside the tube cap (20), a waste liquid pool, a partitioned liquid storage structure (30), and a needle inserting structure (40) are provided from top to bottom respectively; The needle inserting structure (40) is used to pierce the partitioned liquid storage structure (30); the needle inserting structure (40) includes a disc-shaped base (41) and a needle. The needle is fixed on the disc-shaped base (41). When the tube cap (20) approaches the tube body (10), the disc-shaped base (41) is supported by the nozzle of the tube body (10); The opening end of the liquid discharge channel on the tube cap (20) is located below the partitioned liquid storage structure (30), and the other end is located at the inner wall near the cap top of the tube cap (20) to prevent the reaction liquid from flowing back after entering the waste liquid chamber; A liquid discharge channel is provided between the waste liquid pool and the tube body (10). When inverted, the liquid drains from the tube body (10) to the waste liquid pool. When upright, the liquid remains in the waste liquid pool.

2. The sampling tube according to claim 1, characterized in that, The partitioned liquid storage structure (30) is provided with a partition ring (31) and an outer ring (32), which are divided into an inner area and an outer area. A sealing film (33) is provided at the bottom of the partitioned liquid storage structure (30).

3. The sampling tube according to claim 2, characterized in that, The lengths of the needles corresponding to the inner and outer areas are different, which is used to release the liquid in the partitioned liquid storage structure (30) step by step.

4. The sampling tube according to claim 1, characterized in that, The sampling tube body (10) has a structure that is larger at the top and smaller at the bottom, and the bottom is conical, which can be inserted into the reaction hole of a conventional PCR instrument. The diameter of the opening end is 1 cm.

5. The sampling tube according to claim 1, characterized in that, The cross-section of the protruding parts of the first protruding ring (11) and the second protruding ring (12) is semi-circular, and the cross-section of the concave parts of the first concave ring (21) and the second concave ring (22) is semi-circular. The four have the same radius; The center distance between the first protruding ring (11) and the second protruding ring (12) is the same as the center distance between the first concave ring (21) and the second concave ring (22).

6. The sampling tube according to claim 2, characterized in that, The partition ring (31) and the outer ring (32) are concentrically distributed, and the outer side and the top surface of the outer ring (32) are connected to the inner wall and the top surface of the tube cap (20).

7. The sampling tube according to claim 2, characterized in that, The four long hollow needles (42) and one short hollow needle (43) of the needle inserting structure (40) are distributed in a plum blossom shape on the disc-shaped base (41). The one short hollow needle (43) is located at the center; circular holes are provided at the positions of the disc-shaped base (41) corresponding to the needles to facilitate the reaction reagent to flow out after the needles pass through the sealing film (33); The length difference between the four long hollow needles (42) and the short hollow needle (43) is consistent with the center distance between the first protruding ring (11) and the second protruding ring (12).

8. The sampling tube according to claim 1, characterized in that, The edge of the disc-shaped base (41) of the pin structure (40) is recessed inward, with a semi-circular cross-section, and engages with the sealing rubber ring (44).

9. An application method of the sampling tube according to claim 7, characterized in that, The reaction reagent A is contained inside the sampling tube body (10); the reaction reagent B is contained between the partition ring (31) and the outer ring (32) of the partitioned liquid storage structure (30), and the reaction reagent C is contained inside the partition ring (31); The steps are as follows: When the sample is added to the sampling tube, the tube cap (20) is covered. After the reaction, the magnetic beads in the reaction reagent A are adsorbed at the bottom of the tube body (10) by the external magnetic force outside the bottom of the tube body (10). The sampling tube is rotated 180° clockwise as a whole, so that the waste liquid completely enters the waste liquid pool through the drainage channel. Then the sampling tube is rotated 180° clockwise again, and the tube cap (20) is pressed downward until the first protruding ring (11) is completely engaged with the second concave ring (22). The long hollow pin (42) in the pin structure (40) punctures the sealing film (33), and the reaction reagent B is released into the tube body. The external magnet at the bottom of the tube body (10) is removed; after the reaction, the tube body is rotated 90° counterclockwise, and an external magnetic force is applied to the left tube wall to adsorb the magnetic beads; while continuously applying the external magnetic force to the left tube wall and rotating the tube body 90° clockwise, the tube cap (20) is pressed downward until the first protruding ring (11) is completely engaged with the first concave ring (21), and the second protruding ring (12) is completely engaged with the second concave ring (22). The short hollow pin (43) in the pin structure (40) punctures the sealing film (33), and the reaction reagent C is released into the tube body for reaction.

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