A kit for quantitative detection of anti-myeloperoxidase antibody IgG

Abstract

The application provides a kit for completely homogeneous determination of anti-myeloperoxidase antibody IgG quantitative detection and a detection method and application thereof. The kit combines chemiluminescence analysis technology and immunological technology, is a cleaning-free kit, is a one-step method, and can be determined in only 15 min. The kit is simple to operate, can realize high-throughput detection by cooperating with a full-automatic instrument due to no cleaning process, and has a CV within 3%. The kit has high sensitivity, a wide linear range, rapid reaction and simple operation, and particularly, the use of MPO inhibitors can inhibit MPO peroxidase activity, thereby reducing the background, improving the distinction of positive and negative sample values, and having important value in the auxiliary diagnosis of immune-related diseases.

Description

Technical Field

[0001] This invention belongs to the field of in vitro detection technology, and particularly relates to a kit for quantitative detection of antimyeloperoxidase antibody IgG and its application in the biomedical field. Background Technology

[0002] Myeloperoxidase (MPO), also known as peroxidase, is a heme protease with a heme cofactor and a member of the heme peroxidase superfamily. A positive anti-medullary peroxidase antibody test is often a marker for necrotizing vasculitis or idiopathic crescentic glomerulonephritis. This antibody positivity can also be seen in other diseases, such as polyarteritis nodosa, anti-glomerular basement membrane disease, granulomatous polyangiitis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced lupus, and Felty's syndrome. Therefore, anti-medullary peroxidase antibody testing is an antibody detection method; by detecting one's own antibodies, many diseases can be identified or diagnosed.

[0003] Currently, clinical detection of anti-MPO antibody IgG includes indirect immunofluorescence, Western blotting, enzyme immunoassay, and magnetic microparticle chemiluminescence. Indirect immunofluorescence and Western blotting are primarily manual methods and provide qualitative results. Enzyme-linked immunosorbent assay (ELISA) has a long reaction time and provides semi-quantitative results. Magnetic microparticle chemiluminescence, when used with a fully automated chemiluminescence analyzer, allows for quantitative detection; however, due to the two-step washing process, the testing time is long and the coefficient of variation (CV) is relatively large. Summary of the Invention

[0004] In a first aspect, the present invention provides a method for detecting antimyeloperoxidase (MPO) antibody IgG, the method comprising detecting antimyeloperoxidase antibody IgG using MPO antigen treated with an MPO inhibitor, wherein the MPO inhibitor comprises an azide compound and / or a benzoyl hydrazine compound.

[0005] Preferably, the azide compounds include, but are not limited to, sodium azide, lithium azide, potassium azide, rubidium azide, or cesium azide.

[0006] Preferably, the benzoyl hydrazide compounds include, but are not limited to, 4-aminobenzoyl hydrazide, 4-hydroxybenzoyl hydrazide, 4-chlorobenzoyl hydrazide, 4-methoxybenzoyl hydrazide or 3-methoxybenzoyl hydrazide.

[0007] The detection method includes adding an azide compound to the MPO antigen, then labeling the MPO antigen treated with the azide compound with a chemiluminescent label, and then adding a benzoyl hydrazine compound.

[0008] Preferably, the concentration of the azide compound is 0.1-2‰ (W / V), for example 0.1‰, 0.2‰, 0.3‰, 0.4‰, 0.5‰, 1‰, 1.5‰, or 2‰ (W / V).

[0009] Preferably, the concentration of the benzoyl hydrazine compound is 0.1-5‰ (W / V), for example 0.1‰, 0.2‰, 0.3‰, 0.4‰, 0.5‰, 1‰, 1.5‰, 2‰, 3‰, 4‰, 5‰ (W / V).

[0010] Preferably, the chemiluminescent marker is selected from luminol, isoluminol and its derivatives, acridine derivatives, 1,2-dioxane and its derivatives; more preferably, the chemiluminescent marker is selected from acridine derivatives; and more preferably, the acridine derivative is selected from 9,10-dihydroacridine.

[0011] Preferably, the detection method includes:

[0012] The sample was mixed with MPO antigen treated with MPO inhibitor and enzyme-labeled antibody to form a complex, a substrate was added, and the light signal intensity was detected.

[0013] The MPO antigen treated with the MPO inhibitor includes adding 0.1-2‰ (W / V) of an azide compound to the MPO antigen, then labeling the MPO antigen treated with the azide compound with a chemiluminescent label, and then adding 0.1-5‰ (W / V) of a benzoyl hydrazine compound.

[0014] Preferably, the enzyme is selected from one or more combinations of peroxidase, glucose oxidase, pyruvate kinase, glucose-6-phosphate dehydrogenase, luciferase, galactosidase, or alkaline phosphatase. The peroxidase includes glutathione peroxidase, eosinophil peroxidase, thyroid peroxidase, or horseradish peroxidase (HRP). More preferably, the enzyme is horseradish peroxidase, and the enzyme-labeled antibody is a goat anti-human polyclonal antibody labeled with horseradish peroxidase.

[0015] Preferably, the substrate includes hydroxycinnamic acid and / or hydrogen peroxide.

[0016] Preferably, the sample is selected from serum or plasma.

[0017] In a second aspect, the present invention provides a kit for the quantitative detection of anti-myeloperoxidase antibody IgG, the kit comprising an MPO inhibitor, an MPO antigen, and a chemiluminescent label.

[0018] Preferably, the MPO inhibitor includes azide compounds and / or benzoyl hydrazide compounds.

[0019] Preferably, the chemiluminescent label is selected from luminol, isoluminol and its derivatives, acridine derivatives or 1,2-dioxane and its derivatives, and more preferably, the acridine derivative is selected from 9,10-dihydroacridine.

[0020] Preferably, the kit further includes one or more of the following: enzyme-labeled antibody, substrate, anti-MPO antibody IgG calibrator, or anti-MPO antibody IgG quality control.

[0021] Preferably, the enzyme is selected from one or more combinations of peroxidase, glucose oxidase, pyruvate kinase, glucose-6-phosphate dehydrogenase, luciferase, galactosidase or alkaline phosphatase, and the peroxidase includes glutathione peroxidase, eosinophil peroxidase, thyroid peroxidase or horseradish peroxidase. More preferably, the enzyme-labeled antibody is a goat anti-human polyclonal antibody labeled with horseradish peroxidase.

[0022] Preferably, the substrate includes hydroxycinnamic acid and / or hydrogen peroxide.

[0023] Preferably, the anti-MPO antibody IgG calibrator is a positive mixed serum containing anti-MPO antibody IgG, preferably a positive mixed serum containing anti-MPO antibody IgG with a concentration of 0-200 RU / mL, such as 0, 5, 20, 50, 100 and / or 200 RU / mL positive mixed serum containing anti-MPO antibody IgG.

[0024] Preferably, the anti-MPO antibody IgG quality control product is a positive mixed serum containing anti-MPO antibody IgG, and more preferably a positive mixed serum containing anti-MPO antibody IgG with concentrations of 20-100 RU / mL, such as 20 RU / mL, 50 RU / mL, 70 RU / mL, and 100 RU / mL.

[0025] In one specific embodiment, the kit includes an anti-MPO antibody IgG calibrator, an anti-MPO antibody IgG quality control, an anti-MPO antibody IgG reagent 1 (Anti-MPO IgG-R1), an anti-MPO antibody IgG reagent 2 (Anti-MPO IgG-R2), and a substrate, wherein the anti-MPO antibody IgG reagent 1 is an acridine derivative-labeled MPO antigen, and the anti-MPO antibody IgG reagent 2 is an HRP-labeled goat anti-human polyclonal antibody.

[0026] In a third aspect, the present invention provides the application of the above-described reagent kit or the above-described detection method in the diagnosis or auxiliary diagnosis of immune-related diseases.

[0027] Preferably, the application is for the purpose of diagnosing and treating diseases, or for purposes other than diagnosing and treating diseases.

[0028] Preferably, the immune-related diseases are selected from primary vasculitis, primary necrotizing crescentic glomerulonephritis (NCGN), allergic granulomatous vasculitis (CSS), polyarteritis nodosa (PAN), anti-glomerular basement membrane disease (anti-GBM disease), Wegener's granulomatosis (WG), systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), drug-induced lupus (DIL), Sjögren's syndrome (SS), and systemic sclerosis (SSc).

[0029] In a fourth aspect, the present invention provides the application of the above-described kit in the preparation of products for diagnosing or assisting in the diagnosis of immune-related diseases.

[0030] Preferably, the immune-related diseases are selected from primary vasculitis, primary necrotizing crescentic glomerulonephritis (NCGN), granulomatous polyangiitis (CSS), polyarteritis nodosa (PAN), anti-glomerular basement membrane disease (anti-GBM disease), Wegener's granulomatosis (WG), systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), drug-induced lupus (DIL), Sjögren's syndrome (SS), systemic sclerosis (SSc), drug-induced lupus, and Felty's syndrome.

[0031] The “golden carne-1,2-dioxane and its derivatives” described in this invention include, but are not limited to, AMPPD, CSPD, ADP-Star or CDP-Star.

[0032] The “luminol, isoluminol and their derivatives” mentioned in this invention include, but are not limited to, luminol, isoluminol, ABEI, AHEI or ABENH.

[0033] This invention discloses a fully automated homogeneous chemiluminescence technology for the quantitative determination of anti-MPO antibody IgG. It eliminates the need for a washing process, making the reaction process faster and more reliable. It can achieve high-speed and accurate determination of a large number of samples with good CV and simple daily maintenance.

[0034] Based on the principle of homogeneous luminescence technology, the MPO antigen is myeloperoxidase, a peroxidase that interferes with horseradish peroxidase in the kit. The MPO antigen labeled with acridine derivatives results in a very high background in the kit. This invention reduces the background by specially treating the MPO antigen and significantly improves the differentiation between positive and negative samples. The calibrators, quality control samples, reagent 1, reagent 2, and substrate solution in the kit are all optimally formulated for this reaction system, providing strong assurance for the kit's shelf life and detection performance. This invention is a wash-free kit, a one-step method, with a reaction time of only 15 minutes. It is simple to operate; due to the absence of a washing process, combined with fully automated instruments, it can achieve high-throughput detection with a CV of less than 3%.

[0035] The above only summarizes some aspects of the present invention and is not, and should not be considered as limiting the present invention in any way.

[0036] All patents and publications mentioned in this specification are incorporated herein by reference in their entirety. Those skilled in the art will recognize that certain modifications may be made to this invention without departing from its spirit or scope.

[0037] The following embodiments further illustrate the present invention in detail, but should not be construed as limiting the scope of the present invention or the specific methods described herein. Attached Figure Description

[0038] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings, wherein:

[0039] Figure 1 The correlation between the measured values ​​of anti-MPO antibody IgG in the samples was detected using this application and the Euromon ELISA kit.

[0040] Figure 2 : Blank limit evaluation of fitted curve;

[0041] Figure 3 Linear range evaluation curve. Detailed Implementation

[0042] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as a result. However, these embodiments are merely exemplary and do not constitute any limitation on the scope of the present invention. Those skilled in the art should understand that modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but all such modifications and substitutions fall within the protection scope of the present invention.

[0043] Example 1: Chemiluminescent Quantitative Detection Kit for Anti-Myeloperoxidase (MPO) Antibody IgG

[0044] The kit components include anti-MPO antibody IgG calibrator, anti-MPO antibody IgG quality control, anti-MPO antibody IgG reagent, and substrate solution. The formulations of each component are optimal for this reaction system, providing strong assurance for the kit's shelf life and detection performance.

[0045] 1. The main components of the reagent kit are as follows:

[0046] • Anti-MPO antibody IgG calibrator (Anti-MPO IgG-STD): Positive mixed serum containing anti-MPO antibody IgG.

[0047] • Anti-MPO antibody IgG quality control (Anti-MPO IgG-QC): Positive mixed serum containing anti-MPO antibody IgG.

[0048] • Anti-MPO antibody IgG reagent 1 (Anti-MPO IgG-R1): MPO antigen labeled with acridine derivative.

[0049] • Anti-MPO IgG Reagent 2 (Anti-MPO IgG-R2): HRP-labeled goat anti-human polyclonal antibody.

[0050] • Substrate solution: Contains p-hydroxycinnamic acid and hydrogen peroxide

[0051] Among them, the anti-MPO antibody IgG reagent No. 1 requires special treatment of the MPO antigen, including the addition of sodium azide and 4-aminobenzoyl hydrazine.

[0052] 2. Reaction principle of the reagent kit

[0053] This study combines chemiluminescence immunoassay with immunoassay. After mixing reagents 1 and 2, a goat anti-human IgG antibody-anti-MPO antibody IgG-MPO antigen complex is formed. Upon addition of the substrate solution, HRP catalyzes the generation of hydroxyl radicals. The acridine derivative then produces a light signal under the influence of these hydroxyl radicals. Within a certain range, the light signal intensity is directly proportional to the concentration of anti-MPO antibody IgG. The anti-MPO antibody IgG content of the sample can be calculated from the calibration curve using the luminescence intensity.

[0054] 3. The specific preparation steps of the reagent kit are as follows:

[0055] (I) Preparation of anti-MPO antibody IgG calibrators and quality control samples

[0056] I. Preparation of calibrator diluent:

[0057] 1. Measure 800mL of purified water into a container, weigh 3-4g of Na2HPO4·12H2O and add it to the container, then stir and mix thoroughly.

[0058] 2. Weigh 0.2-0.3g of NaH2PO4·2H2O and add it to the container, stirring thoroughly until completely dissolved.

[0059] 3. Weigh 7-10g of NaCl and add it to the container, stirring thoroughly until completely dissolved.

[0060] 4. Adjust the pH; a pH between 7.0 and 8.0 is acceptable.

[0061] 5. Finally, bring the volume to 1L with purified water, filter through a 0.2μm filter, and store at 2-8℃ for later use.

[0062] II. Preparation of anti-MPO antibody IgG calibrator:

[0063] Anti-MPO antibody IgG was prepared to concentrations of 0, 5, 20, 50, 100, and 200 RU / mL using calibrator diluent.

[0064] III. Preparation of anti-MPO antibody IgG quality control material:

[0065] The anti-MPO antibody IgG was prepared to a concentration of 20, 100 RU / mL using calibrator dilution buffer.

[0066] (II) Preparation of Anti-MPO Antibody IgG Reagent No. 1

[0067] 1) Take MPO antigen, add 0.1-2‰ (W / V) sodium azide and treat at 4℃ for 1 hour, then dialyze with PBS overnight.

[0068] 2) Take 1.5 ml centrifuge tubes and add 708.7 μl of 0.05 M pH 8.5 sodium borate buffer, 41.3 μl of acridine derivative labeling reagent dissolved in DMF, and 250 μl of 1 mg / ml treated MPO antigen.

[0069] 3) Vortex and mix 4-5 times, wrap in aluminum foil, and let stand at room temperature for 30 minutes.

[0070] 4) Place the centrifuge tubes at 2-8℃ and mix overnight.

[0071] 5) Dilute the above antigen markers with PTBSTB buffer containing different concentrations (0.1-5‰ (W / V)) of 4-aminobenzoyl hydrazine to a suitable concentration (0.5-2ug / ml).

[0072] (III) Preparation of Anti-MPO Antibody IgG Reagent No. 2

[0073] 1) Use the periodic acid method to label goat anti-human polyclonal antibodies with HRP.

[0074] 2) Dilute the HRP-antibody marker with 1X PBS to an appropriate concentration (0.2-0.5 ug / ml).

[0075] (iv) Preparation of substrate solution

[0076] 1) Measure 800mL of purified water into a container, weigh 8.2g of p-hydroxycinnamic acid and add it to the container, then stir and mix thoroughly.

[0077] 2) Weigh 3.4g of hydrogen peroxide and add it to the container, stirring thoroughly until completely dissolved.

[0078] 3) Finally, bring the volume to 1L with purified water and store at 2-8℃ for later use.

[0079] Example 2: Optimization of reagent kit detection conditions

[0080] Based on the principle of homogeneous luminescence technology, the MPO antigen labeled with acridine derivatives results in a very high background of the kit. This invention reduces the background by specially treating the MPO antigen to inhibit the activity of the peroxidase of the MPO antigen.

[0081] The processing method in this embodiment is the same as that in the preparation of anti-MPO antibody IgG reagent No. 1 in Example 1, except for the addition of sodium azide and 4-aminobenzoyl hydrazine and their concentrations.

[0082] 1. The results of MPO antigen after treatment with sodium azide and labeling are shown in Table 1.

[0083] Table 1 Results of MPO antigen treatment with sodium azide only

[0084]

[0085] Note: The samples were obtained from Beijing 304 Hospital. Negative samples were those that tested negative using the EuroMed ELISA method, and positive samples were those that tested positive using the EuroMed ELISA method.

[0086] 2. The results of adding different concentrations of 4-aminobenzoyl hydrazine to anti-MPO antibody IgG reagent No. 1 dilution (R1-PBSTB) are shown in Table 2.

[0087] Table 2 Results of MPO antigen treatment with 4-aminobenzoylhydrazine only

[0088]

[0089] 3. The results of treating the MPO antigen with 1‰ sodium azide and then labeling it with the anti-MPO antibody IgG reagent No. 1 dilution and then treating it with 4-aminobenzoyl hydrazine are shown in Table 3.

[0090] Table 3 Results of MPO antigen treatment with sodium azide and 4-aminobenzoyl hydrazide

[0091]

[0092] As shown in the table above, the MPO antigen, after being treated with sodium azide and labeled, exhibited generally low luminescence values ​​and poor discrimination. Treatment of anti-MPO antibody IgG reagent dilution 1 with 4-aminobenzoyl hydrazine showed some differentiation between positive and negative samples, but the differentiation was weak. After MPO antigen was treated with sodium azide and labeled, different concentrations of 4-aminobenzoyl hydrazine were added to the R1 reagent dilution, resulting in significant differentiation between positive and negative samples. The best differentiation was observed with the addition of 2‰ 4-aminobenzoyl hydrazine. These results indicate that the combined use of sodium azide and 4-aminobenzoyl hydrazine has a synergistic effect, effectively reducing background and improving the differentiation between positive and negative samples.

[0093] Example 3: Detection of anti-MPO antibody IgG using a kit

[0094] The MPO antigen was treated with 1‰ sodium azide and 2‰ 4-aminobenzoyl hydrazine, and the other steps were the same as in Example 1. The specific detection steps are as follows:

[0095] 1) Add 10 μL of calibrator / quality control sample / sample, 25 μL of reagent 1, and 25 μL of reagent 2, and incubate at 37°C for 15 min.

[0096] 2) Add 50 μL of substrate solution and test the luminescence value.

[0097] 3) Test 20 positive and 20 negative samples.

[0098] The specific steps and results are as follows:

[0099] 1. Obtaining the calibration curve

[0100] Table 4 Calibration Curve Generation Data

[0101]

[0102]

[0103] 2. The specific test data for the samples are shown in Table 5, and the specific test results are as follows: Figure 1 As shown, for the same sample, the results of this application and the Euromon ELISA kit are similar, indicating that this application can effectively and accurately detect anti-MPO antibody IgG. However, the test time of this application can be greatly reduced, with a reaction time of only 15 minutes. The operation is simple and requires no washing process.

[0104] Table 5 Sample Detection Data

[0105]

[0106]

[0107]

[0108] Example 4: Efficacy Verification of the Reagent Kit

[0109] The analytical performance of this kit is evaluated as follows:

[0110] 1) Accuracy Evaluation

[0111] An anti-MPO antibody IgG sample (A) with a concentration of approximately 200 RU / mL (with a concentration deviation of ±20%) was added to a sample B in serum or other appropriate matrix. The volume of A added did not exceed 10% of the total volume (A+B). The recovery rate R was calculated according to formula (1). The recovery rate of this method was in the range of 85-115%. The data are shown in Table 6.

[0112]

[0113] R: Recovery rate;

[0114] V: Volume of sample (A);

[0115] V0: Volume of sample B;

[0116] C: The concentration detected after A+B;

[0117] C0: Detection concentration of sample B;

[0118] C s : Concentration of the standard solution.

[0119] Table 6 Accuracy Evaluation—Adding Recovery Experiment Data

[0120]

[0121]

[0122] 2) Blank Limit Evaluation

[0123] Using a zero-concentration calibrator as the sample, the test was repeated 20 times, and the RLU (relative luminescence value) of the 20 measurements was obtained. The mean (M) and standard deviation (SD) of these 20 measurements were calculated, and the RLU value corresponding to M+2SD was obtained. A two-point regression was performed based on the concentration-RLU values ​​between the zero-concentration calibrator and adjacent calibrators to obtain a linear equation. The RLU value corresponding to M+2SD was substituted into the above equation to obtain the corresponding concentration value, which is the blank limit. The blank limit of this method is not greater than 1 RU / mL. Data are shown in Table 7.

[0124] Table 7 Blank Limit Evaluation—Experimental Data

[0125]

[0126]

[0127] The fitted curve connecting points A and B is shown in the figure. Figure 2

[0128] 3) Linear range evaluation

[0129] High-value samples approaching the upper limit of the linear range (200 RU / mL) were diluted to at least five concentrations at a certain ratio, with low-value samples approaching the lower limit of the linear range. Following the kit instructions, each concentration was tested twice, and the average value was calculated. The average value and dilution ratio were then fitted to a linear relationship using the least squares method, and the linear correlation coefficient r was calculated. The measurement range for this method is [2, 200] RU / mL, and the correlation coefficient r should be ≥0.9900. Data are shown in Table 8. Curves are shown below. Figure 3 .

[0130] Table 8. Evaluation of Linear Range

[0131]

[0132] 4) Repeatability evaluation

[0133] The kits used in this example were used to repeatedly test samples at concentrations of (20±4) RU / mL and (100±20) RU / mL 10 times each. The mean M and standard deviation SD of the 10 measurements were calculated. The coefficient of variation CV was obtained according to the formula CV = SD / M × 100%. The coefficient of variation (CV) of this method is no greater than 8%. See Table 9 for the data.

[0134] CV=SD / M×100%........................(2)

[0135] Where: CV—coefficient of variation; SD—standard deviation of 10 measurements; M—mean of 10 measurements.

[0136] Table 9 Repeatability Evaluation

[0137] Measure serum concentration (RU / mL) Number of measurements Inter-analytical CV (%) 20 10 1.54％ 100 10 2.16％

[0138] 5) Stability evaluation

[0139] Accelerated stability tests were conducted on the kit at 4℃ for 12 months and at 37℃ for 7 days. The results showed that the changes in luminescence intensity of the kit standard, intra-batch and inter-batch precision, and accuracy were all within the normal range, and the kit has a shelf life of up to 12 months.

[0140] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0141] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

[0142] Furthermore, various different embodiments of the present invention can be combined in any way, as long as they do not violate the spirit of the present invention, they should also be regarded as the content disclosed by the present invention.

Claims

1. Use of MPO inhibitor-treated MPO antigen for the preparation of a reagent for the detection of anti-myeloperoxidase antibodies IgG, characterized in that , The MPO inhibitor is sodium azide and 4-amino benzhydrazide; The treatment comprises adding 1-2‰ (W / V) sodium azide to the MPO antigen, then chemiluminescent label the MPO antigen treated by sodium azide, and then adding 1-5‰ (W / V) 4-amino benzhydrazide; The detection method comprises detecting the anti-myeloperoxidase antibody IgG using the MPO antigen treated by the MPO inhibitor, mixing the sample with the MPO antigen treated by the MPO inhibitor and the enzyme-labeled antibody to form a complex, adding a substrate, and detecting the light signal intensity.

2. Use according to claim 1, characterized in that, The chemiluminescent label is selected from luminol, isoluminol and derivatives thereof, acridine derivatives, or adamantane-1,2-dioxetane and derivatives thereof.

3. Use according to claim 1, characterized in that, The enzyme is peroxidase, and the peroxidase comprises glutathione peroxidase, eosinophil peroxidase, thyroid peroxidase, or horseradish peroxidase.

4. Use according to claim 1, characterized in that, The enzyme-labeled antibody is a goat anti-human polyclonal antibody containing horseradish peroxidase.

5. The use according to claim 1, characterized in that, The substrate comprises hydroxycinnamic acid and / or hydrogen peroxide.

6. Use according to claim 1, characterized in that, The sample is selected from serum or plasma. The MPO inhibitor is sodium azide and 4-amino benzhydrazide; The treatment comprises adding 1-2‰ (W / V) sodium azide to the MPO antigen, then chemiluminescent label the MPO antigen treated by sodium azide, and then adding 1-5‰ (W / V) 4-amino benzhydrazide; The detection method comprises detecting the anti-myeloperoxidase antibody IgG using the MPO antigen treated by the MPO inhibitor, mixing the sample with the MPO antigen treated by the MPO inhibitor and the enzyme-labeled antibody to form a complex, adding a substrate, and detecting the light signal intensity. The chemiluminescent label is selected from luminol, isoluminol and derivatives thereof, acridine derivatives, or adamantane-1,2-dioxetane and derivatives thereof. The enzyme is peroxidase, and the peroxidase comprises glutathione peroxidase, eosinophil peroxidase, thyroid peroxidase, or horseradish peroxidase. The enzyme-labeled antibody is a goat anti-human polyclonal antibody containing horseradish peroxidase. The substrate comprises hydroxycinnamic acid and / or hydrogen peroxide. The sample is selected from serum or plasma.

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