Anti-pollution nucleic acid amplification reactor
By designing an anti-pollution nucleic acid amplification reactor, using the sampling port to add the reaction liquid and sample the product, the problems of high nucleic acid amplification detection cost and aerosol pollution in the prior art are solved, and rapid and low-cost nucleic acid detection in scenarios without large laboratories are achieved.
Patent Information
- Application Number
- CN202421863089.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-08-02
- Publication Date
- 2025-06-03
- Estimated Expiration
- 2034-08-02
AI Technical Summary
The existing nucleic acid amplification detection technology has problems of excessive cost while ensuring that nucleic acid amplification products do not cause aerosol pollution to the environment, and it is difficult to widely use in grassroots scenarios without large laboratories.
An anti-pollution nucleic acid amplification reactor is designed, including a reaction tube and a sampler. The reaction tube adds the reaction liquid through the sampling port and takes out the reaction product. After the reaction is completed, the reaction product can be removed without opening the cover, reducing the impact of aerosol contamination.
The product sampling of nucleic acid amplification reaction without opening the cover is realized, reducing the impact of aerosol contamination on the determination of results, simplifying operations, reducing costs, and supporting rapid on-site detection.
Smart Images
Figure CN222935403U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the technical field of laboratory equipment, and particularly provides an anti-pollution nucleic acid amplification reactor, which is particularly applicable to loop-mediated isothermal amplification reaction. Background Technique
[0002] Nucleic acid amplification technology is one of the most commonly used methods in gene diagnosis technology. Currently, methods for nucleic acid amplification include polymerase chain reaction (PCR), nucleic acid sequence-based amplification (NASBA), self-sustained sequence replication (SR), strand displacement amplification (SDA), etc. All of them can rapidly amplify trace specimens, but they have their own disadvantages in terms of specificity, simplicity, temperature and reagent instrument requirements. Since the birth of the PCR method, there have been certain problems in some links, resulting in the nucleic acid detection industry being limited to large laboratories and implementing very strict management measures. Existing nucleic acid detection technologies cannot be widely applied to grass-roots scenarios without large laboratories. Therefore, how to expand the application scenarios of nucleic acid detection technology urgently needs to be solved.
[0003] To solve the problem of pollution by nucleic acid amplification products, currently two main strategies are mainly adopted: 1. Create a closed environment during the detection of nucleic acid amplification products. For example, electrophoresis is carried out in a closed negative-pressure room, and the reaction tube is not opened throughout the process of real-time fluorescence PCR; 2. Use dUTP to replace dTTP in the nucleic acid amplification reaction, and add UNG enzyme to degrade the uracil base in the contaminated product U-DNA, and completely hydrolyze and break the U-DNA strand under high-temperature denaturation conditions to eliminate the amplification caused by contaminated products and ensure the specificity of the amplification result.
[0004] The above first strategy requires strict experimental conditions and expensive experimental facilities and equipment, with too high costs. The second strategy requires temperature-varying operations, has many steps, high overall costs, and there is still a probability of pollution; In summary, the nucleic acid amplification detection technology in the prior art has the problem of too high costs on the premise of ensuring that the nucleic acid amplification products do not cause aerosol pollution to the environment.
[0005] Therefore, in recent years, some new isothermal amplification technologies have developed rapidly, such as loop-mediated isothermal amplification (LAMP), cross-priming isothermal amplification technology (CPA), rolling circle amplification (RCA), transcription-mediated isothermal amplification (TMA), helicase-dependent DNA isothermal amplification, recombinase polymerase amplification (RPA), strand displacement amplification (SDA), etc.; Different from the classical PCR method's dependence on cyclic temperature variation, nucleic acid isothermal amplification technology can rapidly amplify target nucleic acids at a constant temperature, and is an effective method for highly sensitive and highly selective detection of nucleic acids.
[0006] However, all nucleic acid detection methods need to go through many pretreatment steps including biological sample extraction and nucleic acid release. Inevitably, the opening of biological samples is required in these pretreatment steps, which is extremely prone to aerosol contamination and poses a great potential biological safety hazard. Therefore, how to solve the aerosol contamination of nucleic acid amplification and achieve the detection of amplification products without opening the lid is a problem that needs to be solved by those skilled in the art. Summary of the Invention
[0007] To solve the above technical problems, the present utility model provides an anti-pollution nucleic acid amplification reactor.
[0008] The present utility model is implemented as follows: an anti-pollution nucleic acid amplification reactor is provided, which includes a reaction tube and a sampling injector. The reaction tube includes a tube body and a tube cap. The upper end of the tube body is open. The upper part of the tube cap is an upper cover with a diameter larger than the upper end diameter of the tube body. The lower part of the tube cap is a cover body with a diameter slightly smaller than the inner diameter of the upper section of the tube body. A sampling hole is provided in the middle of the side wall of the cover body. A flexible first sealing ring is provided on the outer side of the lower end of the cover body. The outer diameter of the first sealing ring is slightly larger than the inner diameter of the upper section of the tube body. The sampling injector includes a sampling syringe and a sampling needle detachably connected to the sampling syringe. The sampling needle is an arc-shaped needle with a length greater than the distance between the bottom of the tube body and the sampling hole. The outer diameter of the sampling needle is smaller than the diameter of the sampling hole.
[0009] Preferably, the upper section of the tube body is a straight tube section, and the lower section is a tapered tube section with a gradually decreasing diameter.
[0010] More preferably, a flexible second sealing ring is provided on the cover body above the sampling hole. The outer diameter of the second sealing ring is slightly larger than the inner diameter of the upper section of the tube body.
[0011] More preferably, on the symmetric two sides of the outer wall of the upper end of the upper section of the tube body, two connecting rings are provided. A limiting ring is movably connected between the two connecting rings. When the tube cap is completely pressed into the tube body, the distance from one connecting ring, passing above the tube cap, to the other connecting ring is L1. When the tube cap is pulled out a certain distance and the sampling hole is higher than the upper end of the tube body, the distance from one connecting ring, passing above the tube cap, to the other connecting ring is L2. The length of the limiting ring is greater than L1 and less than L2.
[0012] More preferably, a docking ring is provided at the end of the sampling needle connected to the sampling syringe. The docking ring is detachably connected to the sampling syringe.
[0013] More preferably, the volume of the tube body is 0.2 - 1.0 mL; the diameter of the sampling hole is 1 mm.
[0014] More preferably, the sampling needle is a stainless steel needle or a plastic needle, with an outer diameter of 0.5 - 0.7 mm and an inner diameter of 0.4 - 0.6 mm.
[0015] Compared with the prior art, the advantages of the present utility model are as follows:
[0016] 1) The reaction solution can be added and the reaction product can be taken out through the sampling port of the present utility model. After the reaction is completed, the reaction product can be taken out without opening the lid, reducing the influence of aerosol pollution on the result determination.
[0017] 2) The sampling and adding device of the present utility model can be used for on-site detection. The reaction only needs to be heated at 65 °C, and sampling with it can complete the on-site rapid detection.
[0018] 3) The structure of the present utility model is simple. The sampling and adding syringe can quantitatively add samples and take samples, and the operation is convenient. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] The present utility model will be further described in detail below with reference to the drawings and embodiments:
[0020] Figure 1 It is a schematic diagram of the overall structure of the present utility model during sampling and adding (the connection ring and the limit ring are not shown);
[0021] Figure 2 It is a schematic diagram of the reaction tube structure when the tube cap is completely pressed into the tube body;
[0022] Figure 3 It is a schematic diagram of the reaction tube structure when the tube cap is pulled out a certain distance and the sampling and adding hole is higher than the upper end of the tube body. DETAILED DESCRIPTION OF THE EMBODIMENTS
[0023] In order to make the purpose, technical solutions and advantages of the present utility model clearer, the present utility model will be further described in detail below with reference to the drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present utility model and are not used to limit the present utility model.
[0024] Reference Figure 1 、 Figure 2 and Figure 3, the present utility model provides an anti-pollution nucleic acid amplification reactor, which includes a reaction tube and a sampling injector. The reaction tube includes a tube body 1 and a tube cap 2. The upper end of the tube body 1 is open. The upper part of the tube cap 2 is an upper cover 201 with a diameter larger than the upper end diameter of the tube body 1. The lower part of the tube cap 2 is a cover body 202 with a diameter slightly smaller than the inner diameter of the upper section of the tube body 1. A sampling hole 203 is provided in the middle of the side wall of the cover body 202. A flexible first sealing ring 3 is provided on the outer side of the lower end of the cover body 202. The outer diameter of the first sealing ring 3 is slightly larger than the inner diameter of the upper section of the tube body 1. The sampling injector includes a sampling syringe 4 and a sampling needle 5 detachably connected to the sampling syringe 4. The sampling needle 5 is an arc-shaped needle with a length greater than the distance between the bottom of the tube body 1 and the sampling hole 203. The outer diameter of the sampling needle 5 is smaller than the diameter of the sampling hole 203.
[0025] When using the anti-pollution nucleic acid amplification reactor provided by the present utility model for nucleic acid amplification reaction, first press the tube cap 2 completely into the tube body 1, sterilize the sealed reaction tube. After completion, pull the tube cap 2 out a certain distance to make the sampling hole 203 higher than the upper end of the tube body 1, and add the reaction solution sucked by the sampling injector into the tube body 1 through the sampling hole 203. Then press the tube cap 2 completely into the tube body 1 for nucleic acid amplification reaction.
[0026] After the reaction is completed, pull the tube cap 2 out a certain distance to make the sampling hole 203 higher than the upper end of the tube body 1, and insert the sampling needle 5 of the sampling injector through the sampling hole 203 to the lower part of the tube body 1 for sampling. The taken sample can be used for the detection of subsequent reaction products.
[0027] For the convenience of connecting with the tube cap 2 and for facilitating the reaction, the upper section of the tube body 1 is a straight tube section, and the lower section is a tapered tube section with a gradually decreasing diameter.
[0028] In order to prevent pollutants such as aerosols from entering the reaction tube through the sampling hole 203 after the tube cap 2 is completely pressed into the tube body 1, as an improvement, a flexible second sealing ring 6 is provided on the cover body 202 above the sampling hole 203. The outer diameter of the second sealing ring 6 is slightly larger than the inner diameter of the upper section of the tube body 1.
[0029] In order to prevent the tube cap 2 from being pulled out of the tube body 1 due to excessive force when the tube cap 2 is pulled out a certain distance and the height of the sampling hole 203 is higher than the tube body 1, as an improvement of the technical solution, on the outer wall of the upper end of the upper section of the tube body 1, two connecting rings 7 are symmetrically provided on both sides. A limiting ring 8 is movably connected between the two connecting rings 7. Let the distance between one connecting ring 7 passing above the tube cap 2 and then to the other connecting ring 7 be L1 when the tube cap 2 is completely pressed into the tube body 1. Let the distance between one connecting ring 7 passing above the tube cap 2 and then to the other connecting ring 7 be L2 when the tube cap 2 is pulled out a certain distance and the sampling hole 203 is higher than the upper end of the tube body 1. The length of the limiting ring 8 is greater than L1 and less than L2.
[0030] The positions of the connecting ring 7 and the limiting ring 8 are set at the upper end of the straight pipe section, which will not affect the placement position of the reaction tube in the PCR instrument.
[0031] When it is necessary to pull out the tube cap 2 by a certain distance, first rotate the limiting ring 8 above the tube cap 2, and then use hand to pull out the tube cap 2. After the sampling and injection hole 203 is higher than the tube body 1, the limiting ring 8 will hold the tube cap 2 to prevent it from being pulled out of the tube body 1.
[0032] For the convenience of assembly, a docking ring 9 is provided at one end of the sampling and injection needle 5 connected to the sampling and injection syringe 4, and the docking ring 9 is detachably connected to the sampling and injection syringe 4.
[0033] Preferably, the volume of the tube body 1 is 0.2 - 1.0 mL, for example, it can be 0.2 mL, 0.5 mL, 1.0 mL; the diameter of the sampling and injection hole 203 is 1 mm.
[0034] Preferably, the sampling and injection needle 5 is a stainless - steel needle, with an outer diameter of 0.5 - 0.7 mm and an inner diameter of 0.4 - 0.6 mm.
[0035] The embodiments of the present utility model have been described in detail above in conjunction with the accompanying drawings. However, the present utility model is not limited to the above - mentioned embodiments. Within the scope of knowledge possessed by those of ordinary skill in the art, various changes can be made without departing from the purpose of the present utility model.
Claims
1. A contamination-resistant nucleic acid amplification reactor, characterized in that: The invention comprises a reaction tube and a sampler. The reaction tube comprises a tube body (1) and a tube cover (2). The tube body (1) has an opening at the upper end. The upper part of the tube cover (2) is an upper cover (201) having a diameter greater than the diameter of the upper end of the tube body (1). The lower part of the tube cover (2) is a cover body (202) having a diameter slightly smaller than the inner diameter of the upper section of the tube body (1). A sampling hole (203) is provided in the middle of the side wall of the cover body (202). A flexible first sealing ring (3) is provided on the outer side of the lower end of the cover body (202). The outer diameter of the first sealing ring (3) is slightly greater than the inner diameter of the upper section of the tube body (1). The sampler comprises a sampling syringe (4) and a sampling needle (5) detachably connected to the sampling syringe (4). The sampling needle (5) is an arc-shaped needle having a length greater than the distance from the bottom of the tube body (1) to the sampling hole (203). The outer diameter of the sampling needle (5) is smaller than the diameter of the sampling hole (203).
2. The anti-pollution nucleic acid amplification reactor according to claim 1, characterized in that: The upper section of the tube body (1) is a straight tube section, and the lower section is a pointed tube section with a gradually decreasing diameter.
3. The anti-pollution nucleic acid amplification reactor according to claim 1, characterized in that: A flexible second sealing ring (6) is provided on the cover body (202) above the sampling hole (203), and the outer diameter of the second sealing ring (6) is slightly larger than the inner diameter of the upper section of the tube body (1).
4. The anti-pollution nucleic acid amplification reactor according to claim 1, characterized in that: Two connecting rings (7) are symmetrically arranged on both sides of the outer wall at the upper end of the upper section of the tube body (1), and a limiting ring (8) is movably connected between the two connecting rings (7). When the tube cover (2) is completely pressed into the tube body (1), the distance from one connecting ring (7) through the top of the tube cover (2) to the other connecting ring (7) is L1. When the tube cover (2) is pulled out a certain distance and the sampling hole (203) is higher than the upper end of the tube body (1), the distance from one connecting ring (7) through the top of the tube cover (2) to the other connecting ring (7) is L2. The length of the limiting ring (8) is greater than L1 and less than L2.
5. The anti-pollution nucleic acid amplification reactor according to claim 1, characterized in that: A docking ring (9) is provided at one end of the sampling needle (5) connected to the sampling syringe (4), and the docking ring (9) is detachably connected to the sampling syringe (4).
6. The anti-pollution nucleic acid amplification reactor according to claim 1, characterized in that: The volume of the tube body (1) is 0.2-1.0 mL; the diameter of the sampling hole (203) is 1 mm.
7. The anti-pollution nucleic acid amplification reactor according to claim 1, characterized in that: The sampling needle (5) is a stainless steel needle or a plastic needle with an outer diameter of 0.5-0.7 mm and an inner diameter of 0.4-0.6 mm.