Zinc transporter 8 antibody detection kit and preparation method thereof

Abstract

The present invention belongs to the technical field of in vitro diagnosis, and discloses a zinc transporter 8 antibody detection kit and a preparation method thereof. The detection kit comprises a zinc transporter 8 antigen-coated magnetic microparticle solution, a sheep anti-human IgG antibody fluorescent marker solution, a sample diluent, a calibration solution and a chemiluminescent substrate solution. The zinc transporter 8 antigen-coated magnetic microparticles can be further selected as 3-[3-carboxyallylamide]propyltriethoxysilane surface-modified silica magnetic microspheres or carboxyl MQ silicone resin-coated magnetic microspheres. The detection kit of the present invention uses sheep anti-human IgG as a fluorescent marker to replace the zinc transporter 8 antigen, has a high binding rate with a luminescent marker, and detects by forming a zinc transporter 8 antibody-antigen-sheep anti-human IgG fluorescent marker complex, which can significantly improve the detection sensitivity.

Description

Technical Field

[0001] The present invention belongs to the technical field of in vitro diagnosis, and in particular relates to a zinc transporter 8 antibody detection kit and a preparation method thereof. Background Art

[0002] Antibodies to zinc transporter 8 are newly discovered antibodies specific to type 1 diabetes. They, along with autoantibodies such as protein tyrosine phosphatase autoantibodies (IA-2A), insulin autoantibodies (IAA), glutamic acid decarboxylase antibodies (GADA) and islet cell antibodies (ICA), are all pancreatic autoantibodies and are markers of autoimmune damage to pancreatic β cells. They are key indicators for diagnosing autoimmune diabetes, including type 1 diabetes and latent autoimmune diabetes in adults (LADA), and are used to differentiate autoimmune diabetes from type 2 diabetes.

[0003] At present, the commonly used methods for clinical detection of zinc transporter 8 antibodies are radioimmunoassay, enzyme-linked immunosorbent assay and magnetic particle chemiluminescence. The basic principle of radioimmunoassay is to first use radioactive I 125 The recombinant ZnT8 antigen is labeled, and the specific antibody in the serum first binds to the antigen to form an antigen-antibody complex. After adding the secondary antibody, the complex is incubated to form an antigen-antibody-secondary antibody complex. After centrifugation, the radioactivity is detected to determine the content of the specific antibody in the serum. However, this method is relatively complicated to operate and has a high cost. The enzyme-linked immunosorbent assay also has the problems of complex operation and a large number of reagents. The magnetic particle chemiluminescence method uses the principle of the double antigen sandwich method to detect and determine the relevant luminescence value RLU, which has the advantages of simple operation and low cost. For example, patents CN 106226526A and CN 114184793 A both disclose a zinc transporter 8 antibody chemiluminescence immunoassay kit, including a zinc transporter 8 coated solid phase carrier (magnetic particle) working solution, acridine ester labeled purified zinc transporter 8 working solution, zinc transporter 8 antibody calibration product and chemiluminescent substrate solution. However, its fluorescent marker is zinc transporter 8. The molecular structure of zinc transporter 8 antigen is complex and often exists in the form of polymers. The zinc transporter 8 antigen is labeled with a chemiluminescent marker. The activity of zinc transporter 8 antigen is not high. It is necessary to use a reducing agent to reduce the zinc transporter 8 polymer antigen and then perform a labeling reaction to improve its detection sensitivity, but this also increases the processing procedures.

[0004] In addition, in the process of using magnetic particle chemiluminescence technology to detect zinc transporter 8 antibodies, first, the specific zinc transporter 8 antibody in the sample reacts with the zinc transporter 8 antigen coated on the magnetic particles to form an antigen-antibody complex. Then, under the action of the magnetic field, the magnetic particles are adsorbed to the wall of the reaction tube, and the unbound substances are washed away by the cleaning solution. Then, the antibody fluorescent marker is added to react to form an antibody-antigen-fluorescent labeled antibody complex for detection. Therefore, the binding stability of magnetic particles and antigens and the effect of magnetic particle magnetic field separation have a significant impact on the accuracy of the test results. The binding stability with antigens and antibodies can be improved by surface modification of magnetic particles, but the performance of functionally modified magnetic particles obtained by coupling reactions currently needs to be further improved. Summary of the invention

[0005] In view of the shortcomings and deficiencies of the above prior art, the primary purpose of the present invention is to provide a zinc transporter 8 antibody detection kit. In the detection kit of the present invention, goat anti-human IgG-fluorescent marker is used to replace zinc transporter 8 antigen, and the binding rate with the luminescent marker is high. The detection is performed by forming a zinc transporter 8 antibody-antigen-goat anti-human IgG fluorescent marker complex, which can significantly improve the detection sensitivity.

[0006] Another object of the present invention is to provide a method for preparing the above-mentioned zinc transporter 8 antibody detection kit.

[0007] The purpose of the present invention is achieved through the following technical solutions:

[0008] A zinc transporter 8 antibody detection kit comprises a zinc transporter 8 antigen-coated magnetic particle solution, a goat anti-human IgG antibody fluorescent marker solution, a sample diluent, a calibrator solution and a chemiluminescent substrate solution.

[0009] Furthermore, the zinc transporter 8 antigen-coated magnetic microparticle solution comprises zinc transporter 8 antigen-coated magnetic microparticles at a concentration of 1-5 mg / mL and a Tris buffer at a pH of 7-9.

[0010] Furthermore, the goat anti-human IgG antibody fluorescent marker solution includes a goat anti-human IgG antibody acridinium ester marker with a concentration of 0.1-1 mg / mL and a Tris buffer with a pH of 7-9.

[0011] Furthermore, the sample diluent is a Tris buffer with a pH of 7 to 9, containing 0.2% to 1% bovine serum albumin, 0.05% to 0.2% surfactant and 0.05% to 0.2% preservative.

[0012] Furthermore, the calibrator solutions are zinc transporter 8 antibody standard solutions with concentrations of 5AU / mL, 50AU / mL and 400AU / mL, respectively.

[0013] Furthermore, the chemiluminescent substrate solution includes a pre-excitation solution and an excitation solution; the pre-excitation solution is a NaOH solution with a concentration of 0.05 to 0.25 mol / L, and the excitation solution is a H2O2 solution with a concentration of 0.05 to 0.2 mol / L.

[0014] Furthermore, the zinc transporter 8 antigen-coated magnetic particle solution is prepared by the following method:

[0015] (1) adding silica-coated ferroferric oxide magnetic microspheres to a mixed solvent of water and ethanol and stirring and dispersing the mixture uniformly, then adding a silane coupling agent 3-[3-carboxyallylamide]propyltriethoxysilane and stirring and reacting the mixture, filtering, washing, and drying the product to obtain surface functionally modified magnetic microspheres;

[0016] (2) The surface functional modified magnetic microspheres are mixed and incubated with zinc transporter 8 in a buffer solution to obtain a zinc transporter 8 antigen coated magnetic microparticle solution.

[0017] Furthermore, the volume ratio of water to ethanol in the mixed solvent of water and ethanol in step (1) is 1-10:90-99; the amount of the silane coupling agent 3-[3-carboxyallylamide]propyltriethoxysilane added is 2% to 10% of the mass of the silicon dioxide-coated ferroferric oxide magnetic microspheres.

[0018] Furthermore, in step (2), the mass ratio of the surface functional modified magnetic microspheres and the zinc transporter 8 for mixed incubation is 100:5-10.

[0019] The silane coupling agent 3-[3-carboxyallylamide]propyltriethoxysilane used in the present invention contains both amino and carboxyl groups, has good binding force to zinc transporter 8, and has good stability and magnetic separation effect.

[0020] More preferably, the zinc transporter 8 antigen-coated magnetic particle solution is prepared by the following method:

[0021] 1) adding silicate, end-capping agent 1,3-bis(3-carboxypropyl)tetramethyldisiloxane and ferroferric oxide magnetic nanospheres to a mixed solvent of water and ethanol and stirring and mixing them evenly, heating to 60-90° C. under inert atmosphere protection, then dropping an acid catalyst solution to carry out stirring reaction, filtering, washing and drying after the reaction is completed to obtain carboxyl MQ silicone resin-coated magnetic microspheres;

[0022] 2) The carboxyl MQ silicone resin-coated magnetic microspheres and zinc transporter 8 are mixed and incubated in a buffer solution to obtain a zinc transporter 8 antigen-coated magnetic microparticle solution.

[0023] Furthermore, the silicate in step 1) is at least one of sodium silicate, methyl orthosilicate and ethyl orthosilicate; the mass ratio of the silicate, the capping agent 1,3-bis(3-carboxypropyl)tetramethyldisiloxane and the ferrosoferric oxide magnetic nanospheres added is 10:0.5~2:10~40.

[0024] Furthermore, the volume ratio of water to ethanol in the mixed solvent of water and ethanol in step 1) is 4-7:6-3. The solvent can dissolve or partially dissolve the reaction raw materials silicate and end-capping agent, but does not dissolve the reaction product carboxyl MQ silicone resin. The carboxyl MQ silicone resin generated by the reaction is precipitated in situ on the surface of the magnetic nano-microspheres, achieving good coating of the magnetic nano-particles and surface functional group modification in one step. The product can be separated by simple filtration, and the process is simple.

[0025] Furthermore, in step 2), the mass ratio of the carboxyl MQ silicone resin-coated magnetic microspheres and the zinc transporter 8 for mixed incubation is 100:5-10.

[0026] By further coating the magnetic microspheres with carboxyl MQ silicone resin, the present invention has a more uniform carboxyl distribution, higher activity, better binding stability with zinc transporter 8 and magnetic separation effect, compared with the magnetic microspheres with surface functional modification through coupling with silane coupling agent, thereby further improving the detection accuracy.

[0027] The preparation method of the above zinc transporter 8 antibody detection kit comprises the following preparation steps:

[0028] A. diluting the zinc transporter 8 antigen-coated magnetic microparticles with Tris buffer to a concentration of 1 to 5 mg / mL to obtain a zinc transporter 8 antigen-coated magnetic microparticle solution;

[0029] B. Mixing acridinium ester and goat anti-human IgG antibody in a Tris buffer solution, incubating the mixture, and diluting the mixture to a concentration of 0.1 to 1 mg / mL to obtain a goat anti-human IgG antibody fluorescent marker solution;

[0030] C. Add 0.2% to 1% bovine serum albumin, 0.05% to 0.2% surfactant and 0.05% to 0.2% preservative to Tris buffer to obtain a sample diluent;

[0031] D. Using the sample diluent, the zinc transporter 8 antibody standard was diluted to concentrations of 5 AU / mL, 50 AU / mL, and 400 AU / mL, respectively, to obtain calibrator solutions;

[0032] E. Pack the zinc transporter 8 antigen-coated magnetic microparticle solution, goat anti-human IgG antibody fluorescent marker solution, sample diluent, calibrator solution and chemiluminescent substrate solution according to specifications to obtain a zinc transporter 8 antibody detection kit.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] (1) The detection kit of the present invention uses goat anti-human IgG-fluorescent marker to replace zinc transporter 8 antigen, has a high binding rate with the luminescent marker, and forms a zinc transporter 8 antibody-antigen-goat anti-human IgG fluorescent marker complex for detection, which can significantly improve the detection sensitivity. No additional processing of the fluorescent marker is required, and the detection steps are simple and convenient.

[0035] (2) The magnetic microspheres used in the kit of the present invention can be surface-modified with a specific silane coupling agent, 3-[3-carboxyallylamide]propyltriethoxysilane, which contains both amino and carboxyl groups, has good binding force to zinc transporter 8, and has good stability and magnetic separation effect.

[0036] (3) The magnetic microspheres used in the kit of the present invention can further achieve good coating of magnetic nanoparticles with carboxyl MQ silicone resin and surface functional group modification in one step through a specific process, and the carboxyl distribution is more uniform, the activity is higher, the binding stability with zinc transporter 8 and the magnetic separation effect are better, thereby further improving the detection accuracy. DETAILED DESCRIPTION

[0037] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto.

[0038] Example 1

[0039] A zinc transporter 8 antibody detection kit of this embodiment is prepared by the following method:

[0040] A. Dilute the magnetic microparticles coated with zinc transporter 8 antigen with Tris buffer to a concentration of 2 mg / mL to obtain a magnetic microparticle solution coated with zinc transporter 8 antigen.

[0041] B. Mix acridinium ester (AE) and goat anti-human IgG antibody in Tris buffer solution, incubate, and dilute to a concentration of 0.2 mg / mL to obtain goat anti-human IgG-AE marker solution.

[0042] C. Add 0.5% bovine serum albumin, 0.1% surfactant Triton X-100 and 0.1% preservative sodium azide into Tris buffer to obtain a sample diluent.

[0043] D. Use sample diluent to dilute the zinc transporter 8 antibody standard to concentrations of 5 AU / mL, 50 AU / mL and 400 AU / mL, respectively, to obtain calibrator solutions.

[0044] E. Package the reagents according to the following formula: 3.0 mL / bottle of zinc transporter 8 antigen-coated magnetic microparticle solution, 6.5 mL / bottle of goat anti-human IgG antibody fluorescent marker solution, 13 mL / bottle of sample diluent, 0.5 mL / tube of calibrator solution, and chemiluminescent substrate solution (0.1 mol / L NaOH solution, 1.0 mL / bottle; 0.1 mol / L H2O2 solution, 1.0 mL / bottle) to obtain a zinc transporter 8 antibody detection kit.

[0045] The zinc transporter 8 antigen-coated magnetic particles of this embodiment are prepared by the following method:

[0046] (1) 20 g of silica-coated ferroferric oxide magnetic microspheres (commercially available, average particle size 200 nm) were added to 60 g of a mixed solvent of water and ethanol (V 水 :V 乙醇 =5:95) was stirred and dispersed uniformly, and then 1.5 g of silane coupling agent 3-[3-carboxyallylamide]propyltriethoxysilane was added to carry out stirring reaction. The product was filtered, washed and dried to obtain magnetic microspheres with surface functional modification.

[0047] (2) The surface functional modified magnetic microspheres and zinc transporter 8 were mixed and incubated in a Tris buffer solution at a mass ratio of 100:6, and diluted to a concentration of 2 mg / mL to obtain a zinc transporter 8 antigen-coated magnetic microparticle solution.

[0048] The detection kit of this embodiment is used to perform detection by indirect immunoassay using magnetic microparticle chemiluminescence technology (using a fully automatic chemiluminescence immunoassay analyzer, calibrating the measurement system with a calibrator before detection to obtain a calibration curve), and the specific detection steps are as follows:

[0049] The first step is incubation: After adding the sample and sample diluent and automatically diluting, take the pre-diluted sample and magnetic particle reagent. The specific zinc transporter 8 antibody in the sample reacts with the zinc transporter 8 antigen coated on the magnetic particles to form an antigen-antibody complex.

[0050] Cleaning: Under the action of the magnetic field, the magnetic particles are adsorbed to the wall of the reaction tube, and the unbound substances are washed away by the cleaning solution.

[0051] The second step of incubation: adding goat anti-human IgG-AE marker to react with the antigen-antibody complex formed in the first step to form an antibody-antigen-goat anti-human IgG complex.

[0052] Wash again.

[0053] Excitation and reading: Pre-excitation solution and excitation solution are added to the reaction complex, and the chemiluminescent reaction is measured by relative luminescence intensity.

[0054] The content of zinc transporter 8 antibody in the sample is positively correlated with the relative luminescence intensity measured by the optical system of the analyzer.

[0055] The test results are determined using a calibration curve.

[0056] Performance Test:

[0057] 1. Linear range test

[0058] The reagent of this example was used to detect five different concentrations of zinc transporter 8 antibody corporate reference products within the linear range (5.00-400.00 AU / mL). Each corporate reference product was tested three times and the average value of each concentration was calculated. According to formulas (1), (2), and (3), the linear equation y=a+bx (y: the detected value of the reagent, x: the actual value of the corporate reference product) and the correlation coefficient r were calculated.

[0059]

[0060]

[0061]

[0062] Where:

[0063] b-slope of the regression line;

[0064] |a| - absolute value of the intercept of the regression line;

[0065] r - correlation coefficient;

[0066] X i - Actual value of the enterprise reference product;

[0067] Y i - The mean of three repeated measurements of the company's reference product;

[0068] i-1, 2, 3, ..., n;

[0069] n - number of samples measured.

[0070] The results showed that within the detection range of the kit 5.00-400.00AU / mL, the linear correlation coefficient r was 0.9952, which met the performance indicator requirement that within the detection range of the kit 5.00-400.00AU / mL, the linear correlation coefficient r should be no less than 0.9900, and the linear relationship was good.

[0071] 2. Detection limit test

[0072] Five samples with a concentration close to 1.00AU / mL were selected for testing. Each sample was tested 5 times, and the test results were sorted by size. The results showed that the test results were all <10.00AU / mL, and there was no test result below the blank limit (0.80AU / mL). This shows that the detection limit of the kit is not greater than 1.00AU / mL.

[0073] 3. Repeatability Test

[0074] Under repeatability conditions, the normal physiological value (5.32AU / mL) and the abnormal physiological value (38.5AU / mL) of the zinc transporter 8 antibody corporate reference product were measured using the kit, and the measurements were repeated 10 times, and the average values ​​of the measured values ​​were calculated respectively ( X ) and standard deviation (S). The coefficient of variation (CV) was calculated according to formula (4).

[0075] CV=S / X *100%......................................(4);

[0076] The results of repeated measurements showed that the coefficient of variation (CV) of the normal physiological value reference was 1.72%, and the coefficient of variation (CV) of the abnormal physiological value reference was 2.66%, which met the performance index requirement that the coefficient of variation (CV) of repeated measurement results should be no more than 10.0%, and the repeated stability was good.

[0077] 4. Accuracy Test

[0078] The kit of this example was used to measure three different concentrations of zinc transporter 8 antibody corporate reference products within the linear range, namely high (200.00 AU / mL), medium (50.00 mg / L), and low (10.00 mg / L). Each corporate reference product was measured three times, the mean of the sample measurement results was calculated, and the relative deviation (Bias%) was calculated according to formula (5).

[0079] Bias%=( X -X) / X*100%................................(5);

[0080] X - Mean value of sample measurement results;

[0081] X-theoretical value of the measured sample.

[0082] The results showed that the relative deviations of the high, medium and low concentrations of the enterprise reference products were -8.52%, -7.46% and -7.89% respectively, which met the performance index requirement that the relative deviation (bias) should not exceed ±10.0%, and had a high accuracy.

[0083] V. Calibrator Measurement Accuracy Test

[0084] The kit calibrator was used as the sample for testing. After repeated measurement 3 times, the absolute deviation (M) of CAL1 (5AU / mL) was calculated according to formula (6). D ), and the relative deviation (B) of CAL2 (50 AU / mL) and CAL3 (400 AU / mL) was calculated according to formula (7): i ).

[0085] M D =|X i -T|........................(6);

[0086] B i =(X i -T) / T×100%................................(7);

[0087] Where:

[0088] X i - Determine the concentration value;

[0089] T—labeled value of the matching calibration material.

[0090] The CAL1 calibrator was tested 3 times. D The results were 0.4AU / mL, 0.5AU / mL, and 0.6AU / mL respectively; the results of Bi of CAL2 calibrator were -5.67%, -4.53%, and -6.20% respectively; the results of Bi of CAL3 calibrator were -7.82%, -7.06%, and -7.61% respectively. The relative deviation between the results of CAL2 and CAL3 and the labeled value should be within ±10.0%; the absolute deviation of CAL1 should be ≤0.8AU / mL, and the accuracy is relatively high.

[0091] Example 2

[0092] In the zinc transporter 8 antibody detection kit of this embodiment, compared with embodiment 1, the zinc transporter 8 antigen-coated magnetic particle solution is prepared by the following method:

[0093] (1) 10 g of methyl orthosilicate, 1.5 g of end-capping agent 1,3-bis(3-carboxypropyl)tetramethyldisiloxane and 20 g of magnetic ferrosoferric oxide nanoparticles were added to a mixed solvent of 60 g of water and ethanol (V 水 :V 乙醇 =5:5) and stirred to mix evenly, heated to 80±5°C under nitrogen protection, and then added with hydrochloric acid catalyst solution for stirring reaction for 2h, and the stirring speed was maintained at 200-250r / min during the reaction process. After the reaction was completed, the mixture was filtered, washed with water and ethanol in sequence, and dried with air flow to obtain carboxyl MQ silicone resin-coated magnetic microspheres.

[0094] (2) Carboxyl MQ silicone resin-coated magnetic microspheres and zinc transporter 8 were mixed and incubated in a Tris buffer solution at a mass ratio of 100:6, and diluted to a concentration of 2 mg / mL to obtain a zinc transporter 8 antigen-coated magnetic microparticle solution.

[0095] The remaining reagent components and specifications of the detection kit of this embodiment are the same as those of Example 1.

[0096] The performance of the test kit of this embodiment was tested, and the results showed that within the test range of 5.00-400.00AU / mL, the linear correlation coefficient r was 0.9989; the detection limit of the test kit was not greater than 1.00AU / mL; the repeated measurement results showed that the coefficient of variation (CV) of the normal physiological value reference was 1.15%, and the coefficient of variation (CV) of the abnormal physiological value reference was 2.67%. The accuracy test results of the enterprise reference showed that the relative deviation results of the high, medium and low concentration enterprise references were 1.52%, -1.64% and -0.91%, respectively. The accuracy test results of the calibration products showed that the CAL1 calibration product was measured 3 times and the M D The results were 0.1AU / mL, 0.1AU / mL, and 0.1AU / mL, respectively; the accuracy test results of the CAL2 calibrator showed that the Bi results of the three measurements were 0.88%, 1.24%, and 1.06%, respectively; the Bi results of the CAL3 calibrator were -2.07%, -1.85%, and -1.20%, respectively.

[0097] By comparing the results with those of Example 1, it can be seen that the present invention can further improve the linear relationship and significantly improve the detection accuracy by using carboxyl MQ silicone resin to in-situ coat and surface carboxyl modify the magnetic nanoparticles compared to the surface modification by silane coupling agent.

[0098] Comparative Example 1

[0099] Compared with Example 1, this comparative example uses zinc transporter 8 antigen-AE marker instead of goat anti-human IgG-AE marker, and the rest is the same, and detection is performed by forming an antigen-antibody-zinc transporter 8 antigen-AE complex.

[0100] The detection sensitivity of the kits of Example 1 and Comparative Example 1 was tested respectively, and the detection method was as follows: the blank sample was measured 20 times, the mean and standard deviation (SD) of the 20 reactions were calculated, and the corresponding concentration was calculated as the mean value + 2SD, which was the analytical sensitivity of the reagent.

[0101] The test results show that the sensitivity of Example 1 is 0.16 AU / mL, and the sensitivity of Comparative Example 1 is 0.52 AU / mL, which is more than 3 times higher.

[0102] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A zinc transporter 8 antibody detection kit, characterized in that: It includes a zinc transporter 8 antigen-coated magnetic microparticle solution, a goat anti-human IgG antibody fluorescent marker solution, a sample diluent, a calibration solution and a chemiluminescent substrate solution; The zinc transporter 8 antigen-coated magnetic particle solution is prepared by the following method: 1) Silicate, end-capping agent 1,3-bis(3-carboxypropyl)tetramethyldisiloxane and ferroferric oxide magnetic nanospheres are added to a mixed solvent of water and ethanol and stirred to mix evenly, and the temperature is raised to 60-90° C. under the protection of an inert atmosphere, and then an acid catalyst solution is added dropwise to react with stirring. After the reaction is completed, the solution is filtered, washed and dried to obtain carboxyl MQ silicone resin-coated magnetic microspheres; 2) mixing and incubating the carboxyl MQ silicone resin-coated magnetic microspheres and zinc transporter 8 in a buffer solution to obtain a zinc transporter 8 antigen-coated magnetic microparticle solution; In step 1), the silicate is at least one of methyl orthosilicate and ethyl orthosilicate; the silicate, the end-capping agent 1,3-bis(3-carboxypropyl)tetramethyldisiloxane and the ferroferric oxide magnetic nanospheres are added in a mass ratio of 10:0.5-2:10-40; the volume ratio of water to ethanol in the mixed solvent of water and ethanol is 4-7:6-3; The goat anti-human IgG antibody fluorescent marker solution comprises a goat anti-human IgG antibody acridinium ester marker with a concentration of 0.1-1 mg / mL and a Tris buffer with a pH of 7-9.

2. A zinc transporter 8 antibody detection kit according to claim 1, characterized in that: The zinc transporter 8 antigen-coated magnetic particle solution comprises zinc transporter 8 antigen-coated magnetic particles with a concentration of 1-5 mg / mL and a Tris buffer with a pH of 7-9.

3. A zinc transporter 8 antibody detection kit according to claim 1, characterized in that, The sample diluent is a Tris buffer with a pH of 7 to 9, containing 0.2% to 1% bovine serum albumin, 0.05% to 0.2% surfactant and 0.05% to 0.2% preservative; the calibration solution is a zinc transporter 8 antibody standard solution with concentrations of 5AU / mL, 50AU / mL and 400AU / mL, respectively.

4. A zinc transporter 8 antibody detection kit according to claim 1, characterized in that: The chemiluminescent substrate solution includes a pre-excitation solution and an excitation solution; the pre-excitation solution is a NaOH solution with a concentration of 0.05-0.25 mol / L, and the excitation solution is a H2O2 solution with a concentration of 0.05-0.2 mol / L.

5. A zinc transporter 8 antibody detection kit according to claim 1, characterized in that, The mass ratio of the carboxyl MQ silicone resin-coated magnetic microspheres and the zinc transporter 8 mixed for incubation in step 2) is 100:5-10.

6. A method for preparing a zinc transporter 8 antibody detection kit according to any one of claims 1 to 5, characterized in that: The method comprises the following preparation steps: A. diluting the magnetic microparticles coated with zinc transporter 8 antigen with Tris buffer to a concentration of 1-5 mg / mL to obtain a magnetic microparticle solution coated with zinc transporter 8 antigen; B. Mixing acridinium ester and goat anti-human IgG antibody in a Tris buffer solution and incubating, diluting to a concentration of 0.1-1 mg / mL to obtain a goat anti-human IgG antibody fluorescent marker solution; C. Add 0.2% to 1% bovine serum albumin, 0.05% to 0.2% surfactant, and 0.05% to 0.2% preservative to Tris buffer to obtain a sample diluent; D. Using the sample diluent, the zinc transporter 8 antibody standard was diluted to concentrations of 5 AU / mL, 50 AU / mL, and 400 AU / mL, respectively, to obtain calibrator solutions; E. Pack the zinc transporter 8 antigen-coated magnetic microparticle solution, goat anti-human IgG antibody fluorescent marker solution, sample diluent, calibrator solution and chemiluminescent substrate solution according to specifications to obtain a zinc transporter 8 antibody detection kit.

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