Kit for tuberculosis antibody detection

Abstract

The invention provides a kit for tuberculosis antibody detection. The kit comprises a sample diluent, an enzyme conjugate reagent, a chromogenic substrate reagent, a concentrated washing liquid reagent and a detection kit. The detection box comprises a sample loader and a reaction part, a reaction cavity is formed in the reaction part, a test strip is arranged in the reaction cavity, and the test strip is provided with a quality control area and a detection area. A liquid inlet is formed in one side of the reaction cavity; a liquid outlet is formed in one side, far away from the liquid inlet, of the reaction cavity; a second liquid inlet is formed in the top of the sample loader, a second liquid outlet is formed in the side wall of the bottom of the sample loader, the size of the second liquid outlet is matched with that of the liquid inlet, and the sample loader is connected with the reaction part in an inserted mode so that the second liquid outlet can be communicated with the liquid inlet. The detection box is provided with the sample loading device and the reaction part, so that the sample loading and the reaction are spatially isolated, the sample loading is simple and easy to operate, the solution in the sample loading stage or the reaction stage is in a closed space, and the phenomenon of inaccurate detection caused by external pollution of the solution can be effectively prevented.

Description

Technical Field

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

[0002] Tuberculosis is a chronic infectious disease caused by infection with Mycobacterium tuberculosis, which poses a huge threat to people's health and life safety. Mycobacterium tuberculosis may invade various organs of the human body, but mainly invades the lungs, which is called pulmonary tuberculosis. Pulmonary tuberculosis refers to tuberculosis lesions occurring in the lungs, trachea, bronchi and pleura, and is mainly divided into five types: primary pulmonary tuberculosis, hematogenously disseminated pulmonary tuberculosis, secondary pulmonary tuberculosis, tracheal (bronchial) tuberculosis, and tuberculous pleurisy. As the world's largest infectious disease, the number of people infected with tuberculosis reaches 2 billion each year, and the number of patients who die from tuberculosis is no less than 1.5 million. The number of deaths is still higher than any other infectious disease. One of the important reasons for this situation is that tuberculosis is transmitted through droplets, the spread speed is extremely fast, and prevention is very difficult. Therefore, rapid and accurate screening and diagnosis of tuberculosis can win valuable treatment time for patients, so as to provide timely treatment and reduce the occurrence of deaths. At the same time, it can also significantly shorten the time patients carry bacteria, thereby greatly reducing the spread and spread of the disease.

[0003] Traditional acid-fast bacilli smear and Mycobacterium tuberculosis isolation and culture are the "gold standard" for tuberculosis diagnosis, but due to the low sensitivity of smear microscopy and the long time-consuming isolation and culture of tuberculosis bacteria, it is difficult to diagnose tuberculosis early, which cannot meet clinical needs. At present, the commonly used methods for detecting serum tuberculosis antibodies are ELISA method and colloidal gold method, but the kits for tuberculosis antibody detection on the market are all for a single or a few tuberculosis antibodies. The detection range is too small, which is easy to cause missed detection. In addition, the existing kit generally includes a test card, and a sample addition port is opened on the test card, and the sample is 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 loading is also cumbersome and inefficient. In this regard, a kit for tuberculosis antibody detection is proposed, which can be used for tuberculosis antibody detection quickly and accurately, and the detection sample is convenient, the air is isolated, the sample contamination is reduced, and the detection accuracy is improved. Summary of the invention

[0004] The present invention aims to provide a kit for tuberculosis antibody detection, which is convenient for sample loading, highly efficient, capable of multi-index detection and high detection accuracy.

[0005] To achieve the above-mentioned 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 liquid reagent and a detection box. The detection box includes 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 just 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 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 sphere is provided below the through hole. The sphere can move up and down to change the distance between the sphere and the through hole. A columnar body is provided at the bottom of the sampler, and 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 ball is connected to a control rod, and the control rod is connected to a handle provided on the outer side wall of the sample loader, and the ball is controlled to move up and down by driving the handle; the ball is connected to the columnar body through a connecting rod assembly, and when the ball moves away from the through hole, it drives the columnar body close to 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 wall of the sample loader 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 sphere, 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 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 near the side of the through hole through a spring, and the spring is used to reset the first baffle and the second baffle.

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

[0010] Preferably, 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.

[0011] Preferably, 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.

[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 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.

[0014] Preferably, 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; and 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 proposes a kit for tuberculosis antibody detection, the kit comprising various detection reagents and a detection box, by arranging the detection box as a sampler and a reaction part, the sampling and the reaction are spatially isolated, the sampling is simple and easy to operate, and the solution is in a closed space in both the sampling and reaction stages, which can effectively prevent the solution from being contaminated by the outside world and cause inaccurate detection. At the same time, the test strip proposed by the present invention is coated with detection lines of multiple antigens, which can realize semi-quantitative detection of tuberculosis-related specific IgG antibodies or IgM antibodies such as ESAT-6 antigen, CFP-10 antigen, clpB antigen, metE antigen, can antigen, ideR antigen, Rv0232 antigen, nrdB antigen, 65Kdantigen antigen, HBHA antigen, Rv0508 antigen, Rv0678 antigen, Rv0792c antigen, Rv0793 antigen, cysK2 antigen, accD2 antigen, Rv0976c antigen, mprA antigen, PE10 antigen, trxB1 antigen in human serum, and realize combined detection of 20 combined diseases, which is fast, comprehensive and accurate. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The figure is a schematic diagram of the structure of a test kit for tuberculosis antibody detection according to the present invention.

[0017] Figure 2 Schematic diagram of the interior of the kit of the present invention.

[0018] Among them, 1-sampler, 2-reaction part, 3-reaction chamber, 4-test strip, 5-sphere, 6-first rod body, 7-third rod body, 8-second rod body, 9-column, 10-first baffle, 11-second baffle. DETAILED DESCRIPTION

[0019] The embodiments described below are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within 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. A groove is provided 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 ball 5, and the other end is fixedly connected to the first rod body 6. When the control handle moves the ball 5, the first rod body 6 is 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 ball 5 contacts the through hole, the columnar body 9 is separated from the second liquid outlet; when the ball 5 moves down and separates from the through hole, the columnar body 9 gradually moves toward the second liquid outlet until it contacts the second liquid outlet, so that the sampler 1 is isolated from the reaction chamber 3. In this way, when adding solution into the sampler 1, the reaction chamber 3 of the reaction part 2 is sealed, and the passage between the reaction chamber 3 and the sampler 1 is opened after the sampler 1 is isolated from the outside, thereby reducing the probability of internal contamination.

[0023] The second liquid inlet is provided with a first baffle 10 and a second baffle 11, and the first baffle 10 and the second baffle 11 are hinged with the rotation axis at the top of the second liquid inlet, and the first baffle 10 and the second baffle 11 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 10 and the second baffle 11. An isolation film is also provided on the top of the first baffle 10 and the second baffle 11 away from the through hole. When in use, the isolation film is torn off first, and then the reagent bottle is directly inserted into the second liquid inlet. Under the action of force, the first baffle 10 and the second baffle 11 can be directly rotated inward to expose the through hole. After the reagent bottle mouth abuts against the through hole, the ball 5 is controlled to move downward, and the solution can be added to the inside of the sampler 1. Then the reagent bottle is pulled out, the first baffle 10 and the second baffle 11 automatically return to the closed state, the ball 5 is controlled to move up to block the through hole, and the column 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 the solution flows to the reaction chamber 3 under the action of gravity to complete the sample loading. After the solution flows into the reaction chamber 3 and reacts with the test strip 4 for a certain period of time, the liquid outlet is opened to discharge the solution.

[0024] The end of the reaction chamber 3 communicating with the liquid outlet is arc-shaped, that is, the reaction chamber 3 and the liquid outlet have arc-shaped side walls, which is conducive to pouring out the liquid in the reaction chamber 3 to avoid residual liquid in the corners.

[0025] A second sliding block is provided at the bottom of the columnar body 9, a guide rail is provided at the bottom of the sample loader 1, and the second sliding block is slidably connected to the guide rail.

[0026] 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.

[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 prepared according to a conventional preparation method, wherein the 20 tuberculosis antigens of the present invention are prepared into corresponding coating concentrations in the preparation of the spot solution, and then the prepared spot solution is fixed on a porous nitrocellulose membrane in a certain arrangement order to form a plurality of independent detection lines spaced at a certain distance, and then drying, film cutting, assembly and other operations are performed to obtain the test strip.

[0030] 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 color developing substrate reagent is a mixture of BCIP and NBT in a molar ratio of 1:2; and the concentrated washing liquid reagent is a 20-fold concentrated Tris buffer.

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

[0032] S1: The tuberculosis antibody detection kit and the sample to be tested are balanced to room temperature, and then the serum sample to be tested is diluted with the sample diluent according to a preset dilution ratio to obtain a diluted sample to be tested, wherein the dilution ratio of the serum sample to be tested to the sample diluent is 1:31, such as adding 1500ul of the sample diluent to a designated container, and then adding 50ul of the serum sample to be tested to obtain a diluted sample to be tested, and the container port of the designated container needs to match the second liquid inlet of the sample loader; then the concentrated washing solution is diluted at a ratio of 1:20 to obtain a working washing solution;

[0033] S2: tear off the isolation film on the first baffle and the second baffle of the sampler, turn the diluted sample to be tested upside down so that the bottle mouth extends into the second liquid inlet of the sampler, and then push the handle to move the ball downward. When the diluted sample to be tested completely flows into the sampler, push the handle to move the ball upward to block the through hole, and take out the container containing the diluted sample to be tested from the second liquid inlet, and then place the entire test box upright so that the sampler is located above the reaction part, add the diluted sample to be tested into the reaction chamber, and the diluted sample to be tested and the ESAT-6 antigen, CFP-10 antigen, clpB antigen, metE antigen, can antigen, ideR antigen, Rv0232 antigen, nrdB antigen, 65Kd coated on the test strip are reacted. The antigen antigen, HBHA antigen, Rv0508 antigen, Rv0678 antigen, Rv0792c antigen, Rv0793 antigen, cysK2 antigen, accD2 antigen, Rv0976c antigen, mprA antigen, PE10 antigen, and trxB1 antigen are incubated 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 antigen, HBHA antigen, Rv0508 antigen, Rv0678 antigen, Rv0792c antigen, Rv0793 antigen, cysK2 antigen, accD2 antigen, Rv0976c antigen, mprA antigen, PE10 antigen, and trxB1 antigen coated on the test strip are combined with 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 chamber, add the working washing solution in S1 to wash the test strip, the working washing solution is added to the reaction chamber through the sample loader, and the loading operation refers to step S2. The working washing solution stays in the reaction chamber for 3 minutes and then is discharged through the liquid outlet. Repeat this operation for 3 times;

[0035] S4: adding the enzyme conjugate reagent to the reaction chamber through the sample loading device. The loading operation is referred to step S2. The enzyme conjugate and the test strip are incubated at 37°C for 10-20 minutes for reaction. The antigen-antibody conjugate on the test strip reacts with the alkaline phosphatase-labeled anti-human IgG antibody or the 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, add the working washing solution in S1 to wash the test strip, the working washing solution is added to the reaction chamber through the sample loader, and the loading operation refers to step S2. The working washing solution stays in the reaction chamber for 3 minutes and then is discharged through the liquid outlet. Repeat this operation for 3 times;

[0037] S6: Add the color substrate reagent to the reaction chamber through the sample loading device. The sample loading operation is referred to step S2. The color substrate reagent and the test strip are incubated at 37° C. for 10-15 minutes to react. The NBT and BCIP mixture in the color substrate reacts with the antigen-antibody conjugate on the test strip in the reaction chamber.

[0038] S7: Open the liquid outlet, pour out the color substrate reagent in the reaction chamber, add the working washing solution in S1 to wash the test strip, the working washing solution is added to the reaction chamber through the sample loader, and the loading operation refers to step S2. The working washing solution stays in the reaction chamber for 3 minutes and then is discharged through the liquid outlet. Repeat this operation for washing 3 times;

[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 of step S8 can be assisted by an immunoblot analyzer. The test strip is interpreted with the help of the immunoblot analyzer, and the results can be divided into five levels: negative, critical value, 2 times the critical value, 4 times the critical value, and 8 times the critical value, and the output is a quantitative value.

[0041] The above-mentioned test kit for tuberculosis antibody detection was validated.

[0042] Population included in the study: Tuberculosis patients from multiple countries and those with suspected tuberculosis symptoms who need differential diagnosis of tuberculosis;

[0043] Sample size included:

[0044] Plan A: 20 subjects, including 13 patients diagnosed with tuberculosis and 7 non-tuberculosis patients;

[0045] Plan B: 100 subjects, including 47 patients diagnosed with tuberculosis and 53 non-tuberculosis patients;

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

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

[0048] Test samples: blood specimens from subjects;

[0049] Detection method: For each of the four schemes A, B, C, and D, two samples of each serum are prepared. The first sample of serum is tested using the test kit for tuberculosis antibody detection proposed in the present invention (hereinafter referred to as closed detection), and the second sample is tested using the same detection reagent, but the test strip is placed in a container connected to the outside, and various reagents are added in sequence in a direct pouring manner. The specific order of addition can refer to the above-mentioned test method (hereinafter referred to as open detection).

[0050] Test results and analysis:

[0051] For plan A, the error rate of both closed and open tests was 0%;

[0052] For plan B, the accuracy of both closed and open tests was 100%;

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

[0054] For Plan D, the accuracy of closed testing was 100%, while the accuracy of open testing was 99.3%, with 4 detection errors.

[0055] This indicates that when the number of test samples increases, the open detection mode is easily affected by external factors and reduces the detection accuracy, while the closed sampling method and reaction conditions are more conducive to obtaining accurate test data.

[0056] The present invention is described in detail above in conjunction with the embodiments. It should be noted that the various specific technical features described in the above specific embodiments can be combined and changed in any suitable manner without contradiction, and the present invention will not further describe various possible combinations. In addition, other variations and combinations of the various technical features of the present invention should also be regarded as the contents disclosed by the present invention and belong to 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 test box; the test 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 just 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 liquid inlet is provided on the top of the sampler There is a second liquid inlet, 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 are connected by plug-in connection 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 sphere is provided below the through hole, the sphere can move up and down and the position distance of the sphere from the through hole can change, a column is provided at the bottom of the sampler, the vertical cross-section of the column matches the size of the second liquid outlet, and 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.

2. The kit for detecting tuberculosis antibodies according to claim 1, characterized in that: The sphere is connected to a control rod, and the control rod is connected to a handle arranged on the outer side wall of the sample loader, and the sphere is controlled to move up and down by driving the handle; the sphere is connected to the columnar body through a connecting rod assembly, and when the sphere moves away from the through hole, it drives the columnar body to approach the second liquid outlet.

3. The kit for detecting tuberculosis antibodies according to claim 2, 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.

4. 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.

5. 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.

6. 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.

7. 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.

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

9. The kit for detecting tuberculosis antibodies according to claim 7, 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.

10. The kit for detecting tuberculosis antibodies according to claim 7, 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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