A disposable closed integrated nucleic acid amplification and detection device

Abstract

The present invention relates to the field of molecular detection technology, and in particular to a disposable sealed integrated nucleic acid amplification and detection device, comprising a nucleic acid amplification container, an intermediate tube and a lid, wherein the two ends of the intermediate tube are sealed and connected to the nucleic acid amplification container and the lid respectively, and the intermediate tube comprises a first chamber, a second chamber, a test strip and a plastic part. The present invention adopts a structural design in which the nucleic acid amplification container, the intermediate tube and the lid are sealed and connected, and the test strip is arranged inside the intermediate tube. After the nucleic acid amplification is completed, the rapid detection of the result is achieved by simple rotation when the entire detection device is sealed, which not only simplifies the operation process of nucleic acid detection, but also effectively prevents aerosol contamination caused by the amplification product, greatly improving the convenience and safety of nucleic acid detection.

Description

Technical Field

[0001] The present invention relates to the field of molecular detection technology, and in particular to a disposable enclosed nucleic acid amplification and detection integrated device. Background Art

[0002] Molecular detection is a technology that uses molecular biological methods to detect nucleic acid molecular fragments. In order to ensure the sensitivity of diagnosis, nucleic acids are generally amplified, and then the amplified products are detected or the changes in the amplification process are monitored to determine whether the sample contains the target sequence. There are three main methods for detecting amplified products. The first is to separate the amplified products by agarose or polypropylene electrophoresis, and use electrophoresis staining to distinguish the target sequence; the second is to label the primers, associate the nucleic acid amplification process with fluorescence, color, pH or precipitation changes, and achieve the purpose of detection by detecting these changes; the third type is to use immunochromatographic test strips to detect amplified products after labeling the primers or probes, and use the convenient and fast characteristics of immunochromatography to facilitate rapid on-site detection.

[0003] At present, the amplification equipment and detection equipment of molecular detection products are separated. The transfer from amplification to detection process involves the transfer of samples after amplification, which will cause aerosol contamination of amplification products. The Chinese invention patent application with application number CN201810669579.9 provides a nucleic acid integrated detection method and detection reagent tube, which integrates amplification and detection equipment. However, after amplification, the amplification tube needs to be taken out and placed inside the device for operation, which is a cumbersome step. Summary of the invention

[0004] In view of the technical problems existing in the above-mentioned prior art, the present invention provides a disposable sealed integrated nucleic acid amplification and detection device with high sealing performance and low cost.

[0005] In order to achieve the above technical objectives, the present invention adopts the following technical solutions:

[0006] A disposable sealed integrated nucleic acid amplification and detection device comprises a nucleic acid amplification container, an intermediate tube and a cover, wherein both ends of the intermediate tube are sealedly connected to the nucleic acid amplification container and the cover respectively, wherein:

[0007] The nucleic acid amplification container comprises:

[0008] A reaction tank, arranged in the middle of the nucleic acid amplification container;

[0009] The intermediate tube comprises:

[0010] A first chamber is provided in the middle of the intermediate tube, and the first chamber is communicated with both ends of the intermediate tube;

[0011] A second chamber is provided on a side of the first chamber, one end of the second chamber is closed, and the other end of the second chamber is connected to one end of the intermediate tube;

[0012] A test strip is arranged inside the first chamber;

[0013] A plastic piece passes through the first chamber, one end of the plastic piece is provided with a plastic baffle, the test paper is provided above the plastic baffle, and one end of the test paper falls into the reaction tank by rotating the plastic piece;

[0014] The cover comprises:

[0015] A slot is provided in the middle of the cover, the plastic part is connected to the cover via the slot, and the cover is rotated to cause the plastic part to rotate accordingly.

[0016] Furthermore, two ends of the intermediate tube are threadedly connected to the nucleic acid amplification container and the cover respectively.

[0017] Furthermore, the cover is provided with a two-stage rotation structure, which includes a rotation assembly structure and a rotation detection structure. The intermediate tube and the cover are rotationally assembled through the rotation assembly structure. The cover is rotated through the rotation detection structure, so that the plastic part follows the rotation and one end of the test paper falls into the reaction tank.

[0018] Furthermore, the second chamber also includes a sealing gasket and a diluent, the sealing gasket is arranged at one end of the second chamber, the sealing gasket is tightly connected to the second chamber to form a diluent cavity, the diluent is arranged in the diluent cavity, and the plastic baffle is rotated to cut the sealing gasket, and the diluent flows into the reaction tank.

[0019] Furthermore, the test paper is a nucleic acid test strip, and the nucleic acid test strip is any one of a PCR amplification product test strip, a LAMP constant temperature amplification product test strip, an RPA constant temperature amplification product test strip, and a CRISPR reaction product test strip.

[0020] Furthermore, the intermediate tube and the nucleic acid amplification container are made of transparent materials.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The present invention adopts a structural design in which a nucleic acid amplification container, an intermediate tube and a lid are sealed and connected, and a test strip is arranged inside the intermediate tube. After the nucleic acid amplification is completed, the result can be quickly detected by simply rotating the entire detection device in a sealed state, which not only simplifies the operation process of nucleic acid detection, but also effectively prevents aerosol contamination caused by amplification products, greatly improving the convenience and safety of nucleic acid detection;

[0023] 2. The present invention realizes the structural conversion of nucleic acid amplification and result detection through a simple rotation step. This process can be combined with simple automated equipment to achieve more convenient operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a schematic diagram of the structure of the present invention;

[0025] Figure 2 It is a schematic diagram of the explosion structure of the present invention;

[0026] Figure 3 It is a schematic diagram of the structure of the nucleic acid amplification container of the present invention;

[0027] Figure 4 is a schematic diagram of the assembly structure of the intermediate tube and the plastic part in an embodiment of the present invention;

[0028] Figure 5 is a schematic diagram of the assembly structure of the intermediate tube and the plastic part in an embodiment of the present invention;

[0029] Figure 6 is a schematic diagram of the assembly structure of the intermediate tube and the sealing gasket in an embodiment of the present invention;

[0030] Figure 7 is a schematic diagram of the structure of the intermediate tube in an embodiment of the present invention;

[0031] Figure 8 is a schematic structural diagram of a cover in an embodiment of the present invention;

[0032] Fig. 9 is a schematic diagram of the assembly structure of the cover and the plastic part in an embodiment of the present invention;

[0033] Description of the drawings: 1. Nucleic acid amplification container; 2. Middle tube; 3. Cover; 4. Plastic part; 5. Reaction tank; 6. First chamber; 7. Second chamber; 8. Plastic baffle; 9. Card slot; 10. Sealing gasket. DETAILED DESCRIPTION

[0034] The embodiments of the present invention are described in detail below with reference to the accompanying drawings, but the present invention can be implemented in many different ways as defined and covered by the claims.

[0035] Example 1

[0036] 1.1 Device assembly

[0037] like Figure 6 As shown, the middle tube 2 is inverted, and the diluent is added into the second chamber 7 of the middle tube 2 , and then the second chamber 7 is sealed using aluminum foil as a sealing gasket 10 .

[0038] like Figure 4 , Figure 5 , Figure 7 , Figure 8 and Fig. 9 As shown, the cover 3 is rotatably connected to the intermediate tube 2 through the rotating assembly structure. At this time, the direction of the slot 9 of the cover 3 is fixed, and the PCR amplification product detection test strip is loaded into the first chamber 6 of the intermediate tube 2 from the lower end, and then the plastic part 4 is inserted into the first chamber 6 of the intermediate tube 2 from the lower end, contacting and inserting into the slot 9. At this time, the plastic baffle 8 at the lower end of the plastic part 4 seals the PCR amplification product detection test strip in the first chamber 6, completing the assembly of the cover 3, the intermediate tube 2, the plastic part 4, the diluent and the PCR amplification product detection test strip, and stored at room temperature.

[0039] 1.2 PCR amplification

[0040] When performing a PCR reaction, various reagents required for the PCR amplification system are added into the nucleic acid amplification container 1 in the manner of ordinary PCR.

[0041] like Figure 1 , Figure 2 and Figure 3 As shown, the nucleic acid amplification container 1 is threadedly and sealedly connected to the lower end of the intermediate tube 2 and placed in a matching nucleic acid amplification instrument for amplification. After the amplification is completed, the cover 3 is screwed to drive the plastic part 4 to rotate, thereby causing the plastic baffle 8 to rotate and cut the sealing gasket 10, allowing the diluent to flow into the reaction tank 5.

[0042] Continue to twist, the plastic baffle 8 no longer blocks the first chamber 6, so that the lower end of the PCR amplification product detection test strip falls into the amplification tube and contacts the liquid in the reaction tank 5. The liquid begins to migrate on the PCR amplification product detection test strip. After the reaction is completed, the detection result of the PCR amplification product detection test strip can be viewed through the transparent middle tube 2.

[0043] Embodiment 2:

[0044] 2.1 Device assembly

[0045] like Figure 6 As shown, the middle tube 2 is inverted, and the diluent is added into the second chamber 7 of the middle tube 2 , and then the second chamber 7 is sealed using aluminum foil as a sealing gasket 10 .

[0046] like Figure 4 , Figure 5 , Figure 7 , Figure 8 and Fig. 9 As shown, the cover 3 is rotatably connected to the intermediate tube 2 through the rotating assembly structure. At this time, the direction of the slot 9 of the cover 3 is fixed, and the LAMP constant temperature amplification product detection test strip is loaded into the first chamber 6 of the intermediate tube 2 from the lower end, and then the plastic part 4 is inserted into the first chamber 6 of the intermediate tube 2 from the lower end, contacting and inserting into the slot 9. At this time, the plastic baffle 8 at the lower end of the plastic part 4 seals the LAMP constant temperature amplification product detection test strip in the first chamber 6, completing the assembly of the cover 3, the intermediate tube 2, the plastic part 4, the diluent and the LAMP constant temperature amplification product detection test strip, and storing it at room temperature.

[0047] 2.2LAMP isothermal amplification

[0048] According to the LAMP constant temperature amplification method, various reagents required for the amplification system are added to the nucleic acid amplification container 1.

[0049] like Figure 1 , Figure 2 and Figure 3 As shown, the nucleic acid amplification container 1 is threadedly and sealedly connected to the lower end of the intermediate tube 2 and placed in a matching nucleic acid amplification instrument for amplification. After the amplification is completed, the cover 3 is screwed to drive the plastic part 4 to rotate, thereby causing the plastic baffle 8 to rotate and cut the sealing gasket 10, allowing the diluent to flow into the reaction tank 5.

[0050] Continue to twist, the plastic baffle 8 no longer blocks the first chamber 6, so that the lower end of the LAMP constant temperature amplification product detection test strip falls into the amplification tube and contacts the liquid in the reaction tank 5. The liquid begins to migrate on the LAMP constant temperature amplification product detection test strip. After the reaction is completed, the detection result of the LAMP constant temperature amplification product detection test strip can be viewed through the transparent middle tube 2.

[0051] Embodiment 3:

[0052] 3.1 Device assembly

[0053] like Figure 4 , Figure 5 , Figure 7 , Figure 8 and Fig. 9As shown, the cover 3 is rotatably connected to the intermediate tube 2 through the rotating assembly structure. At this time, the direction of the slot 9 of the cover 3 is fixed, and the CRISPR reaction product detection test strip is loaded into the first chamber 6 of the intermediate tube 2 from the lower end, and then the plastic part 4 is inserted into the first chamber 6 of the intermediate tube 2 from the lower end, contacting and inserting into the slot 9. At this time, the plastic baffle 8 at the lower end of the plastic part 4 seals the CRISPR reaction product detection test strip in the first chamber 6, completing the assembly of the cover 3, the intermediate tube 2, the plastic part 4, the diluent and the CRISPR reaction product detection test strip, and stored at room temperature.

[0054] 3.2RPA isothermal amplification

[0055] According to the RPA isothermal amplification method and the CRISPR reaction system, various reagents required are added to the nucleic acid amplification container 1.

[0056] like Figure 1 , Figure 2 and Figure 3 As shown, the nucleic acid amplification container 1 is threadedly and sealedly connected to the lower end of the intermediate tube 2, and is placed in a matching nucleic acid amplification instrument for amplification. After RPA amplification and CRISPR are completed, the cover 3 is twisted to drive the plastic part 4 to rotate, and the plastic baffle 8 no longer blocks the first chamber 6, so that the lower end of the CRISPR reaction product detection test strip falls into the amplification tube and contacts the liquid in the reaction tank 5. The liquid begins to swim on the CRISPR reaction product detection test strip. After the reaction is completed, the detection result of the CRISPR reaction product detection test strip is viewed through the transparent intermediate tube 2.

[0057] It can be seen from the above embodiments that the present invention has a simple structure, low cost, good sealing and simple operation. After the nucleic acid amplification is completed, the results can be quickly detected by simple rotation when the entire detection device is sealed, which not only simplifies the operation process of nucleic acid detection, but also effectively prevents aerosol contamination caused by amplification products, greatly improving the convenience and safety of nucleic acid detection.

[0058] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. Any simple modification or transformation made using the contents of the present invention specification, or direct or indirect application in other related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A disposable enclosed integrated nucleic acid amplification and detection device, characterized in that: The invention comprises a nucleic acid amplification container (1), an intermediate tube (2) and a cover (3), wherein two ends of the intermediate tube (2) are sealedly connected to the nucleic acid amplification container (1) and the cover (3), respectively, wherein: The nucleic acid amplification container (1) comprises: A reaction tank (5) is arranged in the middle of the nucleic acid amplification container (1); The intermediate tube (2) comprises: A first chamber (6) is provided in the middle of the intermediate tube (2), and the first chamber (6) is communicated with both ends of the intermediate tube (2); a second chamber (7) disposed on a side of the first chamber (6), one end of the second chamber (7) being closed, and the other end of the second chamber (7) being connected to one end of the intermediate tube (2); A test strip is arranged inside the first chamber (6); A plastic part (4) passes through the first chamber (6), one end of the plastic part (4) is provided with a plastic baffle (8), the test paper is arranged above the plastic baffle (8), and the plastic part is rotated so that one end of the test paper falls into the reaction tank (5); The cover (3) comprises: A card slot (9) is provided in the middle of the cover (3); the plastic part (4) is connected to the cover (3) via the card slot (9); and the cover (3) is rotated so that the plastic part (4) rotates accordingly; The cover (3) is provided with a two-stage rotation structure, the two-stage rotation structure comprising a rotation assembly structure and a rotation detection structure, the intermediate tube (2) and the cover (3) are rotationally assembled through the rotation assembly structure, the cover is rotated through the rotation detection structure, so that the plastic part (4) rotates accordingly, and one end of the test paper falls into the reaction tank (5); The second chamber (7) further comprises a sealing gasket (10) and a diluent. The sealing gasket (10) is arranged at one end of the second chamber (7). The sealing gasket (10) is located on the side of the plastic baffle (8). The sealing gasket (10) is tightly connected with the second chamber (7) to form a diluent cavity. The diluent is arranged in the diluent cavity. By rotating the plastic part (4), the plastic baffle (8) cuts the sealing gasket (10) and allows the diluent to flow into the reaction tank (5).

2. The disposable enclosed integrated nucleic acid amplification and detection device according to claim 1, characterized in that: Both ends of the intermediate tube (2) are respectively threadedly connected to the nucleic acid amplification container (1) and the cover (3).

3. The disposable enclosed integrated nucleic acid amplification and detection device according to claim 1, characterized in that: The test paper is a nucleic acid test strip.

4. The disposable enclosed integrated nucleic acid amplification and detection device according to claim 3, characterized in that: The nucleic acid detection test strip is any one of a PCR amplification product detection test strip, a LAMP constant temperature amplification product detection test strip, an RPA constant temperature amplification product detection test strip, and a CRISPR reaction product detection test strip.

5. The disposable enclosed integrated nucleic acid amplification and detection device according to claim 1, characterized in that: The intermediate tube (2), the nucleic acid amplification container (1) and the cover (3) are made of transparent materials.

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