A kit for tuberculosis antibody detection

Abstract

The present invention provides a kit for detecting tuberculosis antibodies. The kit includes a sample diluent, an enzyme conjugate reagent, a chromogenic substrate reagent, a concentrated washing solution reagent, and a detection cassette. The detection cassette includes a sampler and a reaction part. A reaction chamber is provided in the reaction part, a test strip is provided in the reaction chamber, and a quality control area and a detection area are provided on the test strip. A liquid inlet is provided on one side of the reaction chamber, and a liquid outlet is provided on the side of the reaction chamber away from the liquid inlet; a second liquid inlet is provided at the top of the sampler, and a second liquid outlet is provided on the side wall at the bottom of the sampler. The second liquid outlet matches the size of the liquid inlet. The sampler and the reaction part are connected by plugging to make the second liquid outlet communicate with the liquid inlet. By setting the detection cassette as a sampler and a reaction part, the sampling and the reaction are spatially isolated. The sampling is simple and easy to operate. Whether it is the sampling or the reaction stage, the solution is in a closed space, which can effectively prevent the phenomenon that the solution is contaminated by the outside world and causes inaccurate detection.

Description

Technical Field

[0001] The present invention relates to the technical field of medical detection, and particularly relates to a kit for detecting tuberculosis antibodies. Background Art

[0002] Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis infection, which poses a great threat to people's physical health and life safety. Mycobacterium tuberculosis may invade various organs of the human body, but mainly invades the lungs, known as pulmonary tuberculosis. Pulmonary tuberculosis refers to tuberculosis lesions occurring in the lungs, trachea, bronchi, and pleura, etc., and is mainly divided into five types: primary pulmonary tuberculosis, hematogenous disseminated pulmonary tuberculosis, secondary pulmonary tuberculosis, tracheal (bronchial) tuberculosis, and tuberculous pleurisy. As the world's number one infectious disease today, the number of people infected with tuberculosis reaches 2 billion every year, and the number of patients who die from tuberculosis is not less than 1.5 million. Its fatality rate is still higher than that of any other infectious disease. One of the important reasons for this situation is that tuberculosis is transmitted through droplets, with a very fast spread speed and extremely difficult prevention. Therefore, rapid and accurate screening and diagnosis of tuberculosis can win precious treatment time for patients, so as to provide timely treatment and reduce the occurrence of death events. At the same time, it can also significantly shorten the time when patients carry the bacteria, thereby greatly reducing the spread and diffusion of the disease.

[0003] Traditional acid-fast bacilli smear and Mycobacterium tuberculosis isolation and culture are the "gold standards" for tuberculosis diagnosis. However, due to the limitations of methodology such as low sensitivity of smear microscopy and long time-consuming for mycobacterium isolation and culture, it is very difficult to diagnose tuberculosis at an early stage and cannot meet the clinical needs. Currently, the commonly used methods for detecting serum tuberculosis antibodies are ELISA method and colloidal gold method. However, the kits for detecting tuberculosis antibodies on the current market only detect single or a few tuberculosis antibodies, with too small a detection range and prone to missed detection. In addition, existing kits generally include a detection card with a sample addition port opened on it, and samples are manually loaded through the sample addition port. During the reaction, the sample addition port is exposed to the air, and there may be a problem of sample contamination. In addition, the operation of manual sample loading is also relatively troublesome and has low efficiency. In view of this, a kit for detecting tuberculosis antibodies is proposed, which can be used for rapid and accurate detection of tuberculosis antibodies, is convenient for sample loading, isolates air, reduces sample contamination, and improves detection accuracy. Summary of the Invention

[0004] The purpose of the present invention is to provide a kit for detecting tuberculosis antibodies, which is convenient for sample loading, has high efficiency, can detect multiple indicators, and has high detection accuracy.

[0005] To achieve the above object, the present invention provides a kit for tuberculosis antibody detection. The kit for tuberculosis antibody detection includes a sample diluent, an enzyme conjugate reagent, a chromogenic substrate reagent, a concentrated washing solution reagent, and a detection cassette. The detection cassette includes a sampler and a reaction part. A reaction cavity is provided in the reaction part. A test strip is provided in the reaction cavity. A transparent window is provided directly above the reaction cavity. The test strip is provided with a quality control area and a detection area. The quality control area is coated with a quality control line of anti-human IgG antibody, and the detection area is coated with detection lines of multiple antigens. A liquid inlet is provided on one side of the reaction cavity, and a liquid outlet is provided on the side of the reaction cavity away from the liquid inlet. The top of the sampler is provided with a second liquid inlet, and the side wall of the bottom of the sampler is provided with a second liquid outlet. The second liquid outlet matches the size of the liquid inlet. The sampler and the reaction part are connected by plugging so that the second liquid outlet communicates with the liquid inlet. A through hole is provided at the bottom of the second liquid inlet, and a spherical ball is provided below the through hole. The spherical ball can move up and down to change the position distance from the through hole. A columnar body is provided at the inner bottom of the sampler. The vertical cross section of the columnar body matches the size of the second liquid outlet. The columnar body is slidably connected to the bottom of the sampler to adjust the distance between one end of the columnar body and the second liquid outlet.

[0006] Preferably, the spherical ball is connected to a control rod, and the control rod is connected to a handle provided on the outer side wall of the sampler. The spherical ball is driven to move up and down by driving the handle; the spherical ball is connected to the columnar body through a connecting rod assembly. When the spherical ball moves away from the through hole, the columnar body is driven to approach the second liquid outlet.

[0007] Preferably, the connecting rod assembly includes a first rod body, a second rod body, and a third rod body. One end of the first rod body is slidably connected to the inner side wall of the sampler up and down. A groove is provided in the first rod body. One end of the second rod body is slidably connected to the groove. The other end of the second rod body is fixedly connected to the top surface of the columnar body. One end of the third rod body is fixedly connected to the spherical ball, and the other end is fixedly connected to the first rod body.

[0008] Preferably, the second liquid inlet is provided with a first baffle and a second baffle. The first baffle and the second baffle are hinged to the top rotation shaft of the second liquid inlet. The sides of the first baffle and the second baffle close to the through hole are both connected to the inner side wall of the corresponding second liquid inlet through springs. The springs are used to reset the first baffle and the second baffle.

[0009] Preferably, the end of the reaction cavity communicating with the liquid outlet is arc-shaped.

[0010] Preferably, a second slider is provided at the bottom of the columnar body, and a guide rail is provided at the bottom of the sampler. The second slider is slidably connected to the guide rail.

[0011] Preferably, the detection area is successively coated along the length direction with a detection line for ESAT-6 antigen, a detection line for CFP-10 antigen, a detection line for clpB antigen, a detection line for metE antigen, a detection line for can antigen, a detection line for ideR antigen, a detection line for Rv0232 antigen, a detection line for nrdB antigen, a detection line for 65Kd antigen, a detection line for HBHA antigen, a detection line for Rv0508 antigen, a detection line for Rv0678 antigen, a detection line for Rv0792c antigen, a detection line for Rv0793 antigen, a detection line for cysK2 antigen, a detection line for accD2 antigen, a detection line for Rv0976c antigen, a detection line for mprA antigen, a detection line for PE10 antigen, and a detection line for trxB1 antigen; the detection lines in the detection area are parallel to each other and the intervals between adjacent detection lines are of equal width.

[0012] Preferably, the test strip is a porous nitrocellulose membrane with a pore size of 0.22 μm.

[0013] Preferably, the interval between adjacent detection lines is 2.0 mm, and the quality control line coated with anti-human IgG antibody is 2.0 mm away from the detection line coated with ESAT-6 antigen.

[0014] Preferably, the sample diluent is a Tris buffer containing 3% (mass concentration) bovine serum albumin; the enzyme conjugate in the enzyme conjugate reagent is a mixture formed by alkaline phosphatase-labeled anti-human IgG antibody and alkaline phosphatase-labeled anti-human IgM antibody; the chromogenic substrate reagent is a mixture of BCIP and NBT with a molar ratio of 1:2; the concentrated washing solution reagent is a 20-fold concentrated Tris buffer.

[0015] The beneficial effects of the present invention are as follows: The present invention provides a kit for tuberculosis antibody detection, which includes various detection reagents and a detection box. By setting the detection box as a sampler and a reaction part, the sampling and reaction are spatially isolated. The sampling is simple and easy to operate. Whether in the sampling or reaction stage, the solution is in a closed space, which can effectively prevent the detection inaccuracy caused by external contamination of the solution. At the same time, the test strip proposed by the present invention is coated with detection lines of multiple antigens, which can semi-quantitatively detect specific IgG antibodies or IgM antibodies related to tuberculosis such as ESAT-6 antigen, CFP-10 antigen, clpB antigen, metE antigen, can antigen, ideR antigen, Rv0232 antigen, nrdB antigen, 65Kd antigen, HBHA antigen, Rv0508 antigen, Rv0678 antigen, Rv0792c antigen, Rv0793 antigen, cysK2 antigen, accD2 antigen, Rv0976c antigen, mprA antigen, PE10 antigen, trxB1 antigen, etc. in human serum, realizing the combined detection of 20 items related to tuberculosis, with rapid, comprehensive and accurate detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 FIG. is a schematic structural diagram of the kit for tuberculosis antibody detection of the present invention.

[0017] Figure 2 FIG. is a schematic internal diagram of the kit of the present invention.

[0018] Wherein, 1 - sampler, 2 - reaction part, 3 - reaction chamber, 4 - test strip, 5 - spherical ball, 6 - first rod, 7 - third rod, 8 - second rod, 9 - columnar body, 10 - first baffle, 11 - second baffle. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0019] The embodiments described below are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0020] The present invention provides a kit for tuberculosis antibody detection, the kit for tuberculosis antibody detection includes a sample diluent, an enzyme conjugate reagent, a color substrate reagent, a concentrated washing liquid reagent and a detection box. The detection box includes a sampler 1 and a reaction part 2, a reaction chamber 3 is provided in the reaction part 2, a test strip 4 is provided in the reaction chamber 3, and a transparent inspection window is provided just above the reaction chamber 3, and the transparent inspection window can facilitate the detection personnel to observe the internal situation of the reaction chamber 3 and observe the changes in the detection line of the test strip 4. The test strip 4 is provided with a quality control area and a detection area, the quality control area is coated with a quality control line of anti-human IgG antibody, and the detection area is coated with a plurality of antigen detection lines. A liquid inlet is provided on one side of the reaction chamber 3, and a liquid outlet is provided on the side of the reaction chamber 3 away from the liquid inlet. A second liquid inlet is provided on the top of the sampler 1, and a second liquid outlet is provided on the bottom side wall of the sampler 1, the second liquid outlet matches the size of the liquid inlet, and the sampler 1 and the reaction part 2 are connected by plugging to connect the second liquid outlet with the liquid inlet. A through hole is provided at the bottom of the second liquid inlet, and a ball 5 is provided below the through hole. The ball 5 can move up and down to change the distance between the ball 5 and the through hole. It should be noted that the diameter of the ball 5 is larger than the diameter of the through hole. When the ball 5 contacts the through hole, the through hole is completely blocked by the ball 5, and the inside of the sampler 1 is isolated from the outside world. When the ball 5 moves down and separates from the through hole, the inside of the sampler 1 is connected to the second liquid inlet. A columnar body 9 is provided at the bottom of the sampler 1. The vertical cross-section of the columnar body 9 matches the size of the second liquid outlet. The columnar body 9 is slidably connected to the bottom of the sampler 1 to adjust the distance between one end of the columnar body 9 and the second liquid outlet.

[0021] The ball 5 is connected to a control rod, and the control rod is connected to a handle provided on the outer wall of the sample loader 1, and the ball 5 is controlled to move up and down by driving the handle. That is, the inspector can control the up and down movement of the ball 5 by turning the handle up and down. The ball 5 is connected to the column 9 through a connecting rod assembly, and when the ball 5 moves away from the through hole, it drives the column 9 to approach the second liquid outlet.

[0022] The connecting rod assembly includes a first rod body 6, a second rod body 8 and a third rod body 7. One end of the first rod body 6 is slidably connected to the inner side wall of the sampler 1 up and down. A groove is formed in the first rod body 6. One end of the second rod body 8 is slidably connected to the groove. The other end of the second rod body 8 is fixedly connected to the top surface of the columnar body 9. One end of the third rod body 7 is fixedly connected to the spherical ball 5, and the other end is fixedly connected to the first rod body 6. When the control handle moves the spherical ball 5, the first rod body 6 will be driven to move up and down through the third rod body 7, thereby driving the second rod body 8 to move. Under the traction of the second rod body 8, the columnar body 9 can be driven to move left and right. When the spherical ball 5 contacts the through hole, the columnar body 9 is separated from the second liquid outlet; when the spherical ball 5 moves down and separates from the through hole, the columnar body 9 gradually moves towards the second liquid outlet until it contacts the second liquid outlet, so that the sampler 1 is separated from the reaction chamber 3. In this way, when adding a solution into the sampler 1, the reaction chamber 3 of the reaction part 2 is sealed. After the sampler 1 is separated from the outside, the passage between the reaction chamber 3 and the sampler 1 is opened, reducing the probability of internal contamination.

[0023] The second liquid inlet is provided with a first baffle 10 and a second baffle 11. The first baffle 10 and the second baffle 11 are hinged to the top rotation shaft of the second liquid inlet. The sides of the first baffle 10 and the second baffle 11 close to the through hole are both connected to the inner side wall of the corresponding second liquid inlet through springs. The springs are used to reset the first baffle 10 and the second baffle 11. An isolation film is also provided at the top of the ends of the first baffle 10 and the second baffle 11 away from the through hole. During use, first tear off the isolation film, and then directly insert the reagent bottle into the second liquid inlet. Under the action of force, the first baffle 10 and the second baffle 11 can be directly rotated inwards to expose the through hole. After the reagent bottle mouth abuts against the through hole, then control the spherical ball 5 to move down, and the solution can be added into the sampler 1. Then pull out the reagent bottle, the first baffle 10 and the second baffle 11 automatically return to the closed state, control the spherical ball 5 to move up to block the through hole, and at the same time the columnar body 9 moves to open the second liquid outlet. When the entire test box is placed vertically, the sampler 1 is located at the top of the reaction part 2, and under the action of gravity, the solution flows into the reaction chamber 3 to complete the sampling. After the solution flows into the reaction chamber 3 and reacts with the test strip 4 for a certain time, opening the liquid outlet can drain the solution.

[0024] One end of the reaction chamber 3 communicating with the liquid outlet is arc-shaped, that is, one end of the reaction chamber 3 and the liquid outlet is an arc-shaped side wall, which is beneficial to pour out the liquid in the reaction chamber 3 to avoid residual liquid in the corner.

[0025] A second slider is provided at the bottom of the columnar body 9, and a guide rail is provided at the bottom of the sampler 1. The second slider is slidably connected to the guide rail.

[0026] The detection area is successively coated with a detection line for ESAT-6 antigen, a detection line for CFP-10 antigen, a detection line for clpB antigen, a detection line for metE antigen, a detection line for can antigen, a detection line for ideR antigen, a detection line for Rv0232 antigen, a detection line for nrdB antigen, a detection line for 65Kd antigen, a detection line for HBHA antigen, a detection line for Rv0508 antigen, a detection line for Rv0678 antigen, a detection line for Rv0792c antigen, a detection line for Rv0793 antigen, a detection line for cysK2 antigen, a detection line for accD2 antigen, a detection line for Rv0976c antigen, a detection line for mprA antigen, a detection line for PE10 antigen, and a detection line for trxB1 antigen along the length direction; the detection lines in the detection area are parallel to each other and the intervals between adjacent detection lines are of equal width.

[0027] The test strip 4 is a porous nitrocellulose membrane with a pore size of 0.22 μm.

[0028] The interval between adjacent detection lines is 2.0 mm, and the interval between the quality control line coated with anti-human IgG antibody and the detection line coated with ESAT-6 antigen is 2.0 mm.

[0029] The preparation method of the test strip 4 is carried out according to the conventional preparation method. Among them, in the preparation of the spotting solution, 20 kinds of tuberculosis antigens of the present invention are formulated into corresponding coating concentrations, and then the prepared spotting solution is fixed on the porous nitrocellulose membrane in a certain arrangement order to form several detection lines with independent intervals at a certain distance, and subsequent operations such as drying, film cutting, and assembly can be carried out to obtain the test strip.

[0030] The sample diluent is a Tris buffer solution containing 3% bovine serum albumin by mass concentration; the enzyme conjugate in the enzyme conjugate reagent is a mixture formed by alkaline phosphatase-labeled anti-human IgG antibody and alkaline phosphatase-labeled anti-human IgM antibody; the chromogenic substrate reagent is a mixture of BCIP and NBT with a molar ratio of 1:2; the concentrated washing solution reagent is a 20-fold concentrated Tris buffer solution.

[0031] The test method of the kit for tuberculosis antibody detection is as follows:

[0032] S1: Equilibrate the kit for tuberculosis antibody detection and the sample to be tested to room temperature. Then dilute the serum sample to be tested with the sample diluent according to a preset dilution ratio to obtain a diluted sample to be tested. Among them, the dilution ratio of the serum sample to be tested to the sample diluent is 1:31. For example, add 1500 ul of the sample diluent into a specified container, and then add 50 ul of the serum sample to be tested to obtain a diluted sample to be tested. The mouth of the specified container needs to match the second liquid inlet of the sampler. Then dilute the concentrated washing solution according to a ratio of 1:20 to obtain the working washing solution;

[0033] S2: Tear off the isolation film on the first baffle and the second baffle of the sampler. Invert the diluted sample to be tested so that the bottle mouth extends into the second liquid inlet of the sampler. Then push the handle to move the ball down. When the diluted sample to be tested completely flows into the sampler, push the handle to move the ball up to block the through hole, and remove the container containing the diluted sample to be tested from the second liquid inlet. Then place the entire test kit upright so that the sampler is above the reaction part, and add the diluted sample to be tested into the reaction cavity. The diluted sample to be tested is incubated with the ESAT-6 antigen, CFP-10 antigen, clpB antigen, metE antigen, can antigen, ideR antigen, Rv0232 antigen, nrdB antigen, 65Kd antigen, HBHA antigen, Rv0508 antigen, Rv0678 antigen, Rv0792c antigen, Rv0793 antigen, cysK2 antigen, accD2 antigen, Rv0976c antigen, mprA antigen, PE10 antigen, trxB1 antigen coated on the test strip at 37 °C for 20 - 30 min; The ESAT-6 antigen, CFP-10 antigen, clpB antigen, metE antigen, can antigen, ideR antigen, Rv0232 antigen, nrdB antigen, 65Kd antigen, HBHA antigen, Rv0508 antigen, Rv0678 antigen, Rv0792c antigen, Rv0793 antigen, cysK2 antigen, accD2 antigen, Rv0976c antigen, mprA antigen, PE10 antigen, trxB1 antigen coated on the test strip bind to the corresponding tuberculosis antibodies in the diluted sample to be tested to form antigen-antibodies;

[0034] S3: Open the liquid outlet, pour out the diluted sample to be tested in the reaction cavity, and add the working washing solution in S1 to wash the test strip. The working washing solution is added to the reaction cavity through the sampler. The sampling operation refers to the steps of S2. The working washing solution stays in the reaction cavity for 3 min and then is discharged through the liquid outlet. Repeat this operation 3 times;

[0035] S4: Add the enzyme conjugate reagent into the reaction chamber through a sampler. Refer to step S2 for the sampling operation. Incubate the enzyme conjugate and the test strip at a constant temperature of 37°C for 10 - 20 min for reaction. The antigen-antibody conjugate on the test strip reacts specifically with the alkaline phosphatase-labeled anti-human IgG antibody or alkaline phosphatase-labeled anti-human IgM antibody in the enzyme conjugate to obtain an antigen-antibody conjugate.

[0036] S5: Open the liquid outlet, pour out the enzyme conjugate reagent in the reaction chamber, and add the working washing solution in S1 to wash the test strip. The working washing solution is added into the reaction chamber through a sampler. Refer to step S2 for the sampling operation. The working washing solution stays in the reaction chamber for 3 min and then is discharged through the liquid outlet. Repeat this operation 3 times for washing.

[0037] S6: Add the chromogenic substrate reagent into the reaction chamber through a sampler. Refer to step S2 for the sampling operation. Incubate the chromogenic substrate reagent and the test strip at a constant temperature of 37°C for 10 - 15 min for reaction. The NBT and BCIP mixture in the chromogenic substrate reacts with the antigen-antibody conjugate on the test strip in the reaction chamber to produce a chromogenic reaction.

[0038] S7: Open the liquid outlet, pour out the chromogenic substrate reagent in the reaction chamber, and add the working washing solution in S1 to wash the test strip. The working washing solution is added into the reaction chamber through a sampler. Refer to step S2 for the sampling operation. The working washing solution stays in the reaction chamber for 3 min and then is discharged through the liquid outlet. Repeat this operation 3 times for washing.

[0039] S8: Observe the color development of the test strip after washing in step S7 and determine the test result.

[0040] Among them, the judgment in step S8 can be assisted by an immunoblot analyzer. With the help of the immunoblot analyzer, the test strip is interpreted. The results can be judged and divided into 5 grades: negative, cut-off value, 2 times the cut-off value, 4 times the cut-off value, and 8 times the cut-off value, and the output is a quantified value.

[0041] Verify the above-mentioned kit detection for tuberculosis antibody detection.

[0042] Study population included: tuberculosis patients from multiple countries and those with suspected tuberculosis symptoms who needed differential diagnosis of tuberculosis.

[0043] Sample size included:

[0044] Protocol A: 20 subjects, including 13 tuberculosis patients and 7 non-tuberculosis patients.

[0045] Protocol B: 100 subjects, including 47 tuberculosis patients and 53 non-tuberculosis patients.

[0046] Protocol C: 300 subjects, including 139 tuberculosis patients and 161 non-tuberculosis patients;

[0047] Protocol D: 600 subjects, including 286 tuberculosis patients and 314 non-tuberculosis patients;

[0048] Test samples: Blood specimens of the subjects;

[0049] Test method: For each of the four protocols A, B, C, and D, two copies of each serum were prepared. The first serum was tested using the kit for tuberculosis antibody detection proposed by the present invention (hereinafter referred to as sealed detection). The second serum was tested using the same test reagents, but the test strip was placed in a container communicating with the outside, and various reagents were added in sequence in a direct pouring manner. The specific addition sequence can refer to the above test method (hereinafter referred to as open detection).

[0050] Test results and analysis:

[0051] For Protocol A, the error rates of both sealed detection and open detection were 0%;

[0052] For Protocol B, the accuracies of both sealed detection and open detection were 100%;

[0053] For Protocol C, the accuracy of sealed detection was 100%, while the accuracy of open detection was 99.7%, with 1 case of detection error;

[0054] For Protocol D, the accuracy of sealed detection was 100%, while the accuracy of open detection was 99.3%, with 4 cases of detection error.

[0055] It shows that when the number of test samples increases, the open detection mode is easily affected by the outside world and the detection accuracy decreases, while the sealed sampling method and reaction conditions are more conducive to obtaining accurate test data.

[0056] The present invention has been described in detail in combination with the embodiments. In addition, it should be noted that, in the above specific embodiments, the various specific technical features described can be combined and changed in any appropriate way without contradiction. The present invention will not describe the various possible combination methods separately. In addition, other variations and combinations according to the technical features of the present invention should also be regarded as the content disclosed by the present invention and fall within the protection scope of the present invention.

Claims

1. A kit for detecting tuberculosis antibodies, characterized in that: The test kit for tuberculosis antibody detection comprises a sample diluent, an enzyme conjugate reagent, a colorimetric substrate reagent, a concentrated washing liquid reagent and a detection box; the detection box comprises a sampler and a reaction part, the reaction part is provided with a reaction chamber, the reaction chamber is provided with a test strip, a transparent window is provided directly above the reaction chamber, the test strip is provided with a quality control area and a detection area, the quality control area is coated with a quality control line of anti-human IgG antibodies, and the detection area is coated with detection lines of multiple antigens; a liquid inlet is provided on one side of the reaction chamber, and a liquid outlet is provided on the side of the reaction chamber away from the liquid inlet; a second liquid inlet is provided on the top of the sampler, and a second liquid outlet is provided on the bottom side wall of the sampler, the second liquid outlet matches the size of the liquid inlet, and the sampler and The reaction part is connected to the second liquid outlet through a plug-in connection; a through hole is provided at the bottom of the second liquid inlet, and a ball is provided below the through hole. The ball can move up and down to change the distance between the ball and the through hole; a column is provided at the bottom of the sampler, and the vertical cross-section of the column matches the size of the second liquid outlet. The column is slidably connected to the bottom of the sampler to adjust the distance between one end of the column and the second liquid outlet; the ball is connected to a control rod, and the control rod is connected to a handle provided on the outer wall of the sampler, and the ball is controlled to move up and down by driving the handle; the ball is connected to the column through a connecting rod assembly, and when the ball moves away from the through hole, it drives the column to approach the second liquid outlet.

2. The kit for detecting tuberculosis antibodies according to claim 1, characterized in that: The connecting rod assembly includes a first rod body, a second rod body and a third rod body, one end of the first rod body is slidably connected to the inner wall of the sample loader up and down, a groove is opened in the first rod body, one end of the second rod body is slidably connected to the groove, the other end of the second rod body is fixedly connected to the top surface of the columnar body, one end of the third rod body is fixedly connected to the sphere, and the other end is fixedly connected to the first rod body.

3. The kit for detecting tuberculosis antibodies according to claim 1, characterized in that: The second liquid inlet is provided with a first baffle and a second baffle, and the first baffle and the second baffle are hinged to the top rotating shaft of the second liquid inlet. The first baffle and the second baffle are connected to the inner wall of the corresponding second liquid inlet through a spring near the side of the through hole, and the spring is used to reset the first baffle and the second baffle.

4. The kit for detecting tuberculosis antibodies according to claim 1, characterized in that: The end of the reaction chamber communicating with the liquid outlet is in an arc shape.

5. The kit for detecting tuberculosis antibodies according to claim 1, characterized in that: A second sliding block is provided at the bottom of the columnar body, a guide rail is provided at the bottom of the sample loader, and the second sliding block is slidably connected to the guide rail.

6. The kit for detecting tuberculosis antibodies according to claim 1, characterized in that: The detection area is coated with detection lines of ESAT-6 antigen, CFP-10 antigen, clpB antigen, metE antigen, can antigen, ideR antigen, Rv0232 antigen, nrdB antigen, 65Kd antigen, HBHA antigen, Rv0508 antigen, Rv0678 antigen, Rv0792c antigen, Rv0793 antigen, cysK2 antigen, accD2 antigen, Rv0976c antigen, mprA antigen, PE10 antigen and trxB1 antigen in sequence along the length direction; the detection lines in the detection area are parallel to each other and the intervals between adjacent detection lines are of equal width.

7. The kit for detecting tuberculosis antibodies according to claim 6, characterized in that: The test strip is a porous nitrocellulose membrane with a pore size of 0.22 μm.

8. The kit for detecting tuberculosis antibodies according to claim 6, characterized in that: The interval between adjacent detection lines is 2.0 mm, and the interval between the quality control line coated with anti-human IgG antibody and the detection line coated with ESAT-6 antigen is 2.0 mm.

9. The kit for detecting tuberculosis antibodies according to claim 6, characterized in that The sample diluent is a Tris buffer with a mass concentration of 3% bovine serum albumin; the enzyme conjugate of the enzyme conjugate reagent is a mixture of alkaline phosphatase-labeled anti-human IgG antibody and alkaline phosphatase-labeled anti-human IgM antibody; the colorimetric substrate reagent is a mixture of BCIP and NBT in a molar ratio of 1:2; the concentrated washing solution reagent is a 20-fold concentrated Tris buffer.

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