WO2023179013A1 - 检测IgA免疫复合物的试剂的应用 - Google Patents

检测IgA免疫复合物的试剂的应用 Download PDF

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WO2023179013A1
WO2023179013A1 PCT/CN2022/127450 CN2022127450W WO2023179013A1 WO 2023179013 A1 WO2023179013 A1 WO 2023179013A1 CN 2022127450 W CN2022127450 W CN 2022127450W WO 2023179013 A1 WO2023179013 A1 WO 2023179013A1
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iga
iga immune
immune complex
molecular probe
product
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饶皑炳
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深圳市陆为生物技术有限公司
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements

Definitions

  • the invention belongs to the field of immune detection, and specifically relates to the application of reagents for detecting IgA immune complexes.
  • IgA human immunoglobulin A
  • Fab antigen-binding fragments
  • Fc 1 crystallized fragment
  • IgA combines with the pathogen to initiate the immune response, and the downstream antibodies IgG and IgM , C3 complement and other molecules of the complement pathway participate to form IgA-IgG, IgA-IgM, IgA-C3, IgA-IgG-IgM, etc., collectively referred to as IgA complexes.
  • these pathogen-bound complexes are generally cleared by the immune system, such as phagocytosis by macrophages.
  • IgA Nephropathy IgA Nephropathy
  • IgAN IgA Nephropathy
  • purpura often called Henoch-Schonlein purpura (HSP), or IgA vasculitis (IgAV).
  • IgA nephropathy is a major category of chronic kidney disease.
  • the initial symptoms of IgA nephropathy are mild, and it takes 3 to 5 years for symptoms to become obvious after kidney damage occurs.
  • the gold standard for diagnosis is renal puncture. Therefore, it is difficult to diagnose IgA nephropathy in the early stages using renal puncture, and specific markers are missing in early diagnosis.
  • the indicators related to nephropathy in the blood routine and urine routine in the laboratory including urine protein, hematuria, creatinine, glomerular filtration rate, serum total IgA, etc., are not very specific for IgA nephropathy; the clinical symptoms of Henoch-Schonlein purpura It is easy to be regarded as an allergy, misleading doctors or patients to check the "allergens" that cause the disease, and performing unnecessary allergic reaction tests.
  • Some researchers have proposed a kit for detecting IgA antibodies, but this kit uses several specific antigens as molecular probes to capture specific, single IgA antibodies. These single IgA antibodies are associated with allergic purpura or IgA nephropathy. The correlation is only for a part of the population allergic to these antigens, and the correlation is missing for most patients. Therefore, the kit uses a single IgA antibody as a general biomarker, and its diagnostic specificity is low.
  • the present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention proposes the application of a reagent for detecting IgA immune complexes, and products prepared using the reagent can specifically detect IgA immune complexes.
  • the application of reagents for detecting IgA immune complexes in the preparation of IgA nephropathy diagnostic products is proposed.
  • the reagents include molecular probes, and the molecular probes include the Fc receptor protein of IgA for specific Binds to the IgA immune complex.
  • the Fc receptor protein of IgA (FCAR or CD89) has a strong specific affinity for the IgA immune complex produced during the immune reaction, but has a weak affinity for a single IgA; the Fc receptor protein of IgA is a transmembrane glycoprotein , present on the surface of myeloid cells, such as neutrophils, monocytes, macrophages and eosinophils, mediates immune responses to pathogens, interacts with IgA opsonizing targets and triggers immune defense processes, including Phagocytosis, antibody-dependent cell-mediated cytotoxicity, and stimulation of release of inflammatory mediators.
  • the molecular probe used in the present invention includes the Fc receptor protein of IgA, which can selectively bind to IgA immune complexes with IgA fragments, thereby concentrating the IgA immune complex, and thereby measuring the concentration of the IgA immune complex. Strong specificity.
  • the IgA Fc receptor protein is a recombinant protein or a natural protein.
  • the molecular probe is a solution or lyophilized powder.
  • the molecular probe lyophilized powder needs to be dissolved in a buffer.
  • the buffer is at least one of phosphate buffer or carbonate buffer.
  • the molecular probe is used to specifically bind to the IgA immune complex in serum and/or plasma to characterize the IgA immune activity index.
  • the IgA immune activity index refers to the concentration of IgA immune complexes in serum/plasma.
  • the product includes any one of a kit, a chip or a test strip.
  • the product is selected from kits.
  • the kit includes molecular probe lyophilized powder, coating buffer, and enzyme plate.
  • the coating buffer is used to specifically bind the IgA by dissolving the molecular probe lyophilized powder to coat the molecular probe on the enzyme plate. immune complexes.
  • the coating buffer is carbonate buffer.
  • the kit further includes a standard, an enzyme-labeled secondary antibody, a chromogenic solution, a stop solution, a washing solution, a diluent and a blocking solution.
  • the kit uses the enzyme labeled on the secondary antibody bound to the IgA immune complex to catalyze the substrate in the chromogenic solution to form colored
  • the product is then measured with a microplate reader to detect the absorbance value of the colored product to characterize the concentration of the IgA immune complex to diagnose IgA nephropathy.
  • the detection method of the kit includes the following steps:
  • the molecular probe is a solution, and the molecular probe solution is directly added to the chip so that the molecular probe is coated on the chip for binding to the IgA immune system. Complex.
  • the molecular probe is a lyophilized powder.
  • the lyophilized powder needs to be dissolved in a coating buffer and coated on the chip for binding to the IgA immune complex.
  • a fluorescently labeled secondary antibody is added to the chip, and the concentration of the IgA immune complex is characterized by detecting fluorescence intensity.
  • the molecular probe solution is arranged on the test paper for binding to the IgA immune complex, and then an enzyme-labeled secondary antibody and a chromogenic solution are added, and a chemiluminescence colorimeter is used to To detect the luminescence intensity, and then characterize the concentration of the IgA immune complex.
  • the application of a reagent for detecting IgA immune complexes in preparing a diagnostic product for Henoch-Schönlein purpura is proposed.
  • the reagent includes a molecular probe, and the molecular probe includes the Fc receptor protein of IgA, For specific binding to the IgA immune complex.
  • the molecular probe is a solution or lyophilized powder.
  • the molecular probe is used to specifically bind to the IgA immune complex in serum and/or plasma to characterize the IgA immune activity index.
  • the product includes any one of a kit, a chip or a test strip.
  • the product is selected from a kit.
  • the kit includes molecular probe lyophilized powder, coating buffer, and enzyme plate.
  • the molecular probe is coated on the chip.
  • the molecular probe is coated on the test paper.
  • a diagnostic product for IgA immune complex-related diseases includes a reagent for detecting IgA immune complexes.
  • the reagent includes a molecular probe, and the molecular probe includes IgA. Fc receptor protein.
  • the IgA immune complex-related disease includes at least one of IgA nephropathy or Henoch-Schonlein purpura.
  • the product includes any one of a kit, a chip or a test strip.
  • the products include products that can determine whether the subject suffers from the IgA immune complex-related disease, products that can determine the risk of the subject suffering from the IgA immune complex-related disease, A product capable of diagnosing whether patients with IgA immune complex-related diseases relapse after cure.
  • the diagnostic product proposed by the present invention can not only diagnose whether a patient suffers from IgA immune complex-related diseases, but also determine the patient's prognosis. This diagnostic product integrates multiple diagnostic functions and has broad clinical application prospects.
  • Figure 1 is a schematic diagram of the detection method of the kit proposed in Embodiment 1 of the present invention.
  • Figure 2 is a diagram showing the results of detecting IgA immune activity index in peripheral blood serum of some children's hospital cases, healthy children and healthy adults in Example 2 of the present invention
  • Figure 3 is a comparison chart of the detection results of the IgA immune activity index of three groups of IgA nephropathy patients treated with three methods in Example 3 of the present invention and the untreated group; among them, the left is the first group of patients who were treated with oral hormones ; The middle is the second group of patients, treated with hormone pulse therapy; the right is the third group of patients, treated with the immunosuppressant cyclophosphamide;
  • Figure 4 shows the detection of IgA immune activity indicators, total IgA and glucose-deficient IgA in serum of patients with IgA nephropathy during renal puncture, the first follow-up visit after treatment, and the second follow-up visit after treatment in Example 3 of the present invention.
  • the result chart ; among them, each line represents the detection index of a patient at different time points. The left is the detection of IgA immune activity index, the middle is the detection of total IgA in serum, and the right is the detection of glucose-deficient IgA. detection;
  • Figure 5 is a diagram illustrating the diagnostic effect of IgA nephropathy by detecting serum IgA immune activity index in Example 4 of the present invention, specifically drawing a subject operating curve;
  • Figure 6 is a graph showing the results of detecting IgA immune activity index in peripheral blood serum of child cases and healthy people in Example 5 of the present invention.
  • Figure 7 is a diagram illustrating the diagnostic effect of allergic purpura by detecting serum IgA immune activity index in Example 5 of the present invention, specifically drawing a subject operating curve;
  • Figure 8 is a diagram illustrating the diagnostic effect of allergic purpura by detecting serum total IgA in Example 5 of the present invention, specifically drawing a subject operating curve.
  • test methods used in the examples are conventional methods; unless otherwise stated, the materials and reagents used are commercially available reagents and materials.
  • This embodiment provides a reagent for detecting IgA immune complexes (characterizing the IgA immune activity index by detecting the concentration of IgA immune complexes.
  • the IgA immune activity index refers to the concentration of IgA immune complexes in serum/plasma).
  • the kit includes a molecular probe named HQP001, which is the Fc receptor protein of IgA (FCAR or CD89).
  • the above kit includes: molecular probe HQP001 lyophilized powder, coating buffer, enzyme plate, standard, enzyme-labeled secondary antibody, chromogenic solution, stop solution, washing solution, diluent and blocking solution.
  • molecular probe HQP001 lyophilized powder is recombinant FCAR protein lyophilized powder
  • the standard material is commercial human serum IgA;
  • the enzyme-labeled secondary antibody is horseradish peroxidase (HRP)-labeled mouse anti-human IgA;
  • the chromogenic liquid includes chromogenic liquid A—3,3’,5,5’-tetramethylbenzidine (TMB) and chromogenic liquid B—hydrogen peroxide solution;
  • the stop solution is 10% sulfuric acid
  • the washing solution is 0.15M phosphate buffer containing 0.05% Tween-20;
  • the diluent is washing solution + 1 ⁇ BSA;
  • the blocking solution is washing solution + 5% BSA.
  • Figure 1 shows a schematic diagram of adding it to one well of the 96-well plate), cover it with a sealing film, and place it in a 4°C environment overnight; Shake off the solution in the well, wash the enzyme plate with washing liquid several times, add blocking solution to the plate, cover it with a sealing film, place it in a 4°C environment overnight, wash the plate once with washing liquid, and dry it.
  • the kit proposed in Example 1 was used to detect the IgA immune activity index in the peripheral blood serum of patients diagnosed by renal puncture (IgA nephropathy) to verify the renal puncture diagnosis results.
  • Figure 2 shows the IgA immune activity index in the serum of 12 children's hospital IgA nephropathy patients (diagnosed by renal puncture), 19 healthy children, and 13 healthy adults. Collect peripheral blood from the above three types of people, extract serum, and then use the kit proposed in Example 1 to perform detection according to the detection method proposed in Example 1. The results are shown in Figure 2.
  • the IgA immune activity index is higher than The threshold line 16 is positive, and the threshold line lower than 16 is negative.
  • the positive quality control product is commercial human serum IgA with a concentration of 250ng/mL, of which IgA immune complexes account for about 1 to 2%, and the negative quality control product is.
  • the control products are water and reaction solution.
  • the negative quality control products and positive quality control products for the IgA immune activity index can be improved in the following ways: large-scale testing of the IgA immune activity index of healthy people to establish a normal basic range: 0 ⁇ mean + standard deviation; weak activity High activity (i.e. IgA immune activity index is low): average + standard deviation ⁇ average + 2* standard deviation; strong activity (i.e. IgA immune activity index is high): > average + 2* standard deviation, etc.
  • Example 1 the kit proposed in Example 1 was used to evaluate the medication effect on renal puncture IgA nephropathy patients.
  • the IgA immune activity index directly reflects the inhibitory effect of IgA immunity during treatment, without time lag, and can be used as a complementary indicator of treatment effect; on the other hand, the IgA immune activity index based on peripheral blood serum is simple to operate and the detection cost is low. , through blood testing in multiple time periods, it can establish the time changes of the patient's IgA immune activity with the treatment process, which can help doctors track the ups and downs of IgA immune activity in the patient's body in real time, and grasp the immediate evidence of disease control and treatment effect. Dynamic trends.
  • peripheral blood can be collected before renal puncture, serum can be extracted, and the IgA immune activity index baseline of the case can be established according to the operating procedure of the IgA immune activity index kit in Example 1. After renal puncture is confirmed, peripheral blood is collected for IgA immune activity index testing at time intervals, such as the first day, second day, third day, seventh day, thirtieth day, and sixtieth day of treatment.
  • the ninetieth day, and so on collect peripheral blood for IgA immune activity index detection; for those who need two or more courses of treatment, on the first, second, and third days after the start of each new course of treatment, On the third day, the seventh day, the thirtieth day, the sixtieth day, the ninetieth day, and so on, peripheral blood was collected for IgA immune activity index testing.
  • Those with sufficient financial conditions can increase the frequency of testing as appropriate, while those with limited financial conditions can reduce the frequency of testing as appropriate; after treatment, peripheral blood should be collected every one month or every three months to detect the IgA immune activity index.
  • the time dynamic curve of the IgA immune activity index is thus depicted.
  • This example conducted an early clinical study and collected 174 patients with IgA nephropathy confirmed by puncture. According to the patient's physical indicators, the above-mentioned patients were divided into four groups. One group of 80 patients was not treated, and the other three groups of patients were treated with different treatments. For treatment, 45 patients in the first group were treated with oral hormones, 17 patients in the second group were treated with hormone pulses, and 32 patients in the third group were treated with the immunosuppressant cyclophosphamide. During the puncture diagnosis, the peripheral blood of the three groups of patients was collected once, and the serum was extracted.
  • the serum IgA immune activity index of the three groups of patients was detected using the kit and detection method in Example 1, and the detection values were recorded; After the patients in the three groups received corresponding treatment, they were visited at least once every 3 months. During the visit, the peripheral blood of the three groups of patients was collected and the IgA immune activity index was detected. At the same time, the serum IgA immune activity index of the patients in the untreated group was measured. Tests were also performed and the test values were recorded; statistics were made on the IgA immune activity index test values of the untreated group and the three groups of patients after treatment, and the results are shown in Figure 3. The first follow-up visit of the three treatment groups was compared with the untreated group.
  • the left side shows the situation of the first group of patients after treatment, using oral hormone therapy; the middle shows the situation of the second group of patients after treatment, using hormone therapy. Impact therapy; the right shows the situation of the third group of patients after treatment, treated with the immunosuppressant cyclophosphamide.
  • the population average of the peripheral blood IgA immune activity index after at least 3 months of treatment with the three regimens dropped by more than 50% compared with the untreated group, and there was statistical significance, indicating that the three treatment regimens It has a significant effect on controlling IgA immune response.
  • each patient was tracked sequentially at three time points: the time of renal puncture, the first follow-up visit after treatment, and the second follow-up visit after treatment.
  • kit and detection method in Example 1 to detect the IgA immune activity index as the baseline
  • ELISA human serum/plasma total IgA kit
  • Gd-IgA1 galactose-deficient IgA1
  • KM55 galactose-deficient IgA1
  • KM55 glucose-deficient IgA
  • the left is the detection of IgA immune activity index
  • the middle is the detection of total IgA in serum
  • the right is the detection of glucose-deficient IgA.
  • Figure 4 left shows that the IgA immune activity index continues to decline during the treatment process, although the comparison between the first return visit and the baseline is statistically significant, indicating that the condition is improving during the treatment process; while the overall IgA and glucose-deficient IgA do not show the condition. Improvement, the overall IgA also showed a downward trend, but the first return visit and the second return visit were not statistically significant compared with the baseline; sugar-deficiency IgA showed an upward trend, which may be due to the detection technology of sugar-deficiency IgA of unreliability.
  • the IgA immune activity index also showed correlation with routine urine abnormality indicators and clinical pathology tests.
  • People with a high immune activity index have an average glomerular filtration rate (eGFR) that is 10% lower than those with a low index; for the Oxford classification of pathological diagnosis after renal puncture, people with a high IgA immune activity index have intracapillary hyperplasia ( The ratio of E1) or the severity of tubular atrophy/interstitial fibrosis (T1/2) was more than twice that of people with low index, indicating that the higher the IgA immune activity index in patients with IgA nephropathy, the more severe the renal damage.
  • eGFR average glomerular filtration rate
  • T1/2 tubular atrophy/interstitial fibrosis
  • This example uses the kit proposed in Example 1 to evaluate the risk of IgA nephropathy in patients with abnormal blood routine, urine routine or renal function indicators.
  • IgA nephropathy is a nephritis in which IgA immune complexes are deposited in different tissues of the kidney, causing local immune reactions. It may involve glomeruli, tubulointerstitial and renal blood vessels, and 80% of people are young adults (20-35 years old). In IgA nephropathy In the early stages of the disease, weak abnormalities in routine blood or urine indicators or kidney function indicators are easily ignored by doctors and patients. For patients diagnosed through renal puncture in the future, their kidney damage can last for 3 to 5 years before showing obvious symptoms.
  • This example uses the kit and detection method proposed in Example 1 to comparatively study the serum IgA immune activity index of 108 IgA nephropathy patients (IgAN) and 63 healthy people (Healthy Control, HC).
  • the t-test shows that two people There is a significant difference in the IgA immune activity index of the group.
  • the corresponding receiver operating curve (ROC) diagram is shown in Figure 5.
  • the area under the curve (AUC) is 0.89.
  • the optimal threshold line corresponds to a specificity of 75% and a sensitivity of 89%, indicating that it is of reference value to diagnose the risk of IgA nephropathy by detecting serum IgA immune activity index.
  • the IgA immune activity index can more accurately assess the risk of IgA nephropathy in patients with abnormal blood or urine routine or renal function indicators.
  • a high IgA immune activity index can immediately attract the attention of doctors and patients and provide early warning of IgA nephropathy. The high risk allows patients to start health management and try to delay kidney damage.
  • the kit proposed in Example 1 was used to diagnose whether the patient suffered from Henoch-Schonlein purpura.
  • Increased serum IgA is a characteristic of Henoch-Schonlein purpura, but increased IgA is not specific to purpura because many other infectious diseases can also cause IgA to rise. Since Henoch-Schonlein purpura is an inflammatory injury caused by IgA complexes in blood vessels, the serum IgA immune activity index shows high specificity for Henoch-Schonlein purpura.
  • This example uses the kit in Example 1 to comparatively study the serum IgA immune activity index of 157 children with Henoch-Schonlein purpura (HSPN) and 33 healthy people (HC, including children and adults).
  • HSPN Henoch-Schonlein purpura
  • HC Humidity
  • the average IgA immune activity index is 18.82, 9.15 respectively.
  • HSPN is almost twice that of HC, and the p-value of t-test is 2E-23. The statistical significance of the difference is strong.
  • the results are shown in Figure 6.
  • the horizontal line between 10 and 20 is the maximum value of the IgA immune activity index of healthy people, which is set as the threshold line.
  • Those with IgA immune activity index higher than the threshold line are positive samples and positive quality control products.
  • It is commercial human serum IgA with a concentration of 250ng/mL (of which IgA immune complex accounts for 1 to 2%), and the negative quality control products are water and reaction solution. It can be seen that the IgA immune activity index of most children with allergic purpura exceeds that of normal people, and most of them are 2 times or more than normal people.
  • the receiver operating curve (ROC) diagram of the comparative test is shown in Figure 7.
  • the area under the curve (AUC) is 0.94, the specificity corresponding to the optimal threshold value is 85%, and the sensitivity is 85%, indicating that by detecting serum IgA
  • the immune activity index is valuable for diagnosing allergic purpura.

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Abstract

检测IgA免疫复合物的试剂的应用。试剂包括分子探针，分子探针包括IgA的Fc受体蛋白，用于特异性结合IgA免疫复合物，进而检测IgA免疫复合物的浓度，为IgA免疫复合物相关疾病如IgA肾病、过敏性紫癜提供诊断依据，准确度高；诊断产品包括能检测IgA免疫复合物的试剂，不仅能诊断患者是否患有IgA免疫复合物相关疾病，还能判断患者的预后，这种诊断产品集多种诊断功能于一体，在临床上具有广泛的应用前景。

Description

检测IgA免疫复合物的试剂的应用 技术领域
本发明属于免疫检测领域，具体涉及检测IgA免疫复合物的试剂的应用。
背景技术
在人体自身免疫反应过程中，人体免疫球蛋白A(IgA，包括2个抗原结合片段(Fab)和1个结晶片段(Fc))参与时，IgA与病原体结合启动免疫反应，下游抗体IgG与IgM、补体通路的C3补体等分子的参与形成IgA-IgG、IgA-IgM、IgA-C3、IgA-IgG-IgM等，统称IgA复合物。在正常免疫反应过程中，这些结合了病原体的复合物一般被免疫系统清除，如被巨噬细胞吞噬等。IgA免疫反应异常时，IgA复合物无法被免疫系统有效清除，使其在血液循环系统及泌尿系统中长期滞留，从而导致两种疾病：IgA复合物在肾脏的沉积与滞留导致IgA肾病(IgA Nehphropathy，IgAN)；IgA复合物在血管的沉积导致紫癜，通常称为过敏性紫癜(Henoch-Schonlein purpura，HSP)，或者IgA血管炎(IgAV)。
慢性肾病为十分常见的慢性病，IgA肾病为慢性肾病的一大类，IgA肾病发病初期症状轻微，到肾损伤形成后的症状明显阶段需要3～5年，其诊断金标准为肾穿刺。因此，使用肾穿刺在IgA肾病发病初期很难确诊，而在早期诊断中缺失特异性标志物。目前检验科的血常规及尿常规中与肾病相关的指标，包括尿蛋白、尿血、肌酐酶、肾小球滤过率、血清总IgA等，对于IgA肾病的特异性不高；过敏性紫癜临床上容易被认为是过敏，误导医生或病人去检查诱发疾病的“过敏原”，而进行不必要的过敏反应检测。有研究者提出检测IgA抗体的试剂盒，但是这种试剂盒是以特定的几种抗原作为分子探针，来捕获特定、单一的IgA抗体，这些单一的IgA抗体与过敏性紫癜或IgA肾病的关联只针对这些抗原过敏的一部分人群，对大部分病人缺失关联性，因此该试剂盒以单一的IgA抗体作为一般性的生物标志物，诊断的特异性偏低。
因此，有必要提出一种特异性检测IgA免疫复合物的产品。
发明内容
本发明旨在至少解决上述现有技术中存在的技术问题之一。为此，本发明提出了检测IgA 免疫复合物的试剂的应用，利用该试剂制备的产品能够特异性检测IgA免疫复合物。
根据本发明的一个方面，提出了检测IgA免疫复合物的试剂在制备IgA肾病诊断产品中的应用，所述试剂包括分子探针，所述分子探针包括IgA的Fc受体蛋白，用于特异性结合所述IgA免疫复合物。
根据本发明的一种优选的实施方式，至少具有以下有益效果：
IgA的Fc受体蛋白(FCAR或CD89)对免疫反应过程中产生的IgA免疫复合物有强烈的特异性亲和力，而对单一IgA的亲和力较弱；IgA中Fc的受体蛋白为跨膜糖蛋白，存在于髓系细胞，如中性粒细胞、单核细胞、巨噬细胞和嗜酸性粒细胞表面，介导对病原体的免疫反应，它与IgA调理靶点相互作用并触发免疫防御过程，包括吞噬作用、抗体依赖性细胞介导的细胞毒性和刺激炎症介质的释放。本发明采用的分子探针包括IgA的Fc受体蛋白，可以选择性地结合带有IgA片段的IgA免疫复合物，即可对IgA免疫复合物进行浓缩，进而可以测定IgA免疫复合物的浓度，特异性强。
在本发明的一些实施方式中，所述IgA的Fc受体蛋白为重组蛋白或天然蛋白。
在本发明的一些实施方式中，所述分子探针为溶液或冻干粉。
在本发明的一些实施方式中，分子探针冻干粉需用缓冲液溶解。
在本发明的一些实施方式中，所述缓冲液为磷酸盐缓冲液或碳酸盐缓冲液中的至少一种。
在本发明的一些优选的实施方式中，所述分子探针用于特异性结合血清和/或血浆中的所述IgA免疫复合物，以表征IgA免疫活动性指数。
所述IgA免疫活动性指数是指血清/血浆中的IgA免疫复合物的浓度。
在本发明的一些实施方式中，所述产品包括试剂盒、芯片或试纸中的任一种。
在本发明的一些优选的实施方式中，所述产品选自试剂盒。
在本发明的一些实施方式中，所述试剂盒包括分子探针冻干粉、包被缓冲液、酶标板。
在本发明的一些实施方式中，所述包被缓冲液通过溶解所述分子探针冻干粉以将所述分子探针包被于所述酶标板上，用于特异性结合所述IgA免疫复合物。
在本发明的一些实施方式中，所述包被缓冲液选取碳酸盐缓冲液。
在本发明的一些实施方式中，所述试剂盒还包括标准品、酶标记的二抗、显色液、终止液、洗涤液、稀释液和封闭液。
在本发明的一些优选的实施方式中，所述试剂盒通过与所述IgA免疫复合物结合的所述二抗上标记的所述酶对所述显色液中的底物进行催化，形成有色产物，进而在酶标仪下检测所述有色产物的吸光值，来表征所述IgA免疫复合物的浓度，以对所述IgA肾病进行诊断。
在本发明的一些实施方式中，所述试剂盒的检测方法，包括以下步骤：
S1：将分子探针冻干粉溶解于包被缓冲液中，得分子探针溶液；
S2：将所述分子探针溶液加入酶标板中，覆盖封口膜，置于4℃环境中过夜；用洗涤液洗涤所述酶标板，向所述酶标板中加入封闭液，覆盖所述封口膜，置于4℃环境中过夜，用洗涤液洗涤所述酶标板，干燥得包被有所述分子探针的酶标板；
S3：用稀释液稀释样本得到样本稀释液，然后将所述样本稀释液、标准品梯度稀释液、阴性对照液分别加入所述包被有分子探针的酶标板中，覆盖所述封口膜进行孵育，孵育结束后用洗涤液洗所述酶标板；将酶标记的二抗加入所述酶标板中孵育，洗涤所述酶标板；将显色液加入所述酶标板中，室温避光显色，之后加入终止液，将所述酶标板置于酶标仪中检测吸光值。
在本发明的一些实施方式中，所述分子探针为溶液，直接将分子探针溶液加入所述芯片中，使所述分子探针包被于所述芯片上，用于结合所述IgA免疫复合物。
在本发明的一些实施方式中，所述分子探针为冻干粉，所述冻干粉需溶解于包被缓冲液中包被于所述芯片上，用于结合所述IgA免疫复合物。
在本发明的一些实施方式中，向所述芯片中加入荧光标记的二抗，通过检测荧光强度来表征所述IgA免疫复合物的浓度。
在本发明的一些实施方式中，将分子探针溶液配置于所述试纸上，用于结合所述IgA免疫复合物，然后再加入酶标记的二抗和显色液，通过化学发光显色仪来检测发光强度，进而表征所述IgA免疫复合物的浓度。
根据本发明的第二个方面，提出了检测IgA免疫复合物的试剂在制备过敏性紫癜诊断产品中的应用，所述试剂包括分子探针，所述分子探针包括IgA的Fc受体蛋白，用于特异性结 合所述IgA免疫复合物。
在本发明的一些实施方式中，所述分子探针为溶液或冻干粉。
在本发明的一些优选的实施方式中，所述分子探针用于特异性结合血清和/或血浆中的所述IgA免疫复合物，以表征IgA免疫活动性指数。
在本发明的一些实施方式中，所述产品包括试剂盒、芯片或试纸中的任一种。
在本发明的一些优选的实施方式种，所述产品选自试剂盒。
在本发明的一些实施方式中，所述试剂盒包括分子探针冻干粉、包被缓冲液、酶标板。
在本发明的一些实施方式中，所述分子探针包被于所述芯片上。
在本发明的一些实施方式中，所述分子探针包被于所述试纸上。
根据本发明的第三个方面，提出了一种IgA免疫复合物相关疾病的诊断产品，所述产品包括检测IgA免疫复合物的试剂，所述试剂包括分子探针，所述分子探针包括IgA的Fc受体蛋白。
在本发明的一些实施方式中，所述IgA免疫复合物相关疾病包括IgA肾病或过敏性紫癜中的至少一种。
在本发明的一些实施方式中，所述产品包括试剂盒、芯片或试纸中的任一种。
在本发明的一些实施方式种，所述产品包括能够判断受试者是否患有所述IgA免疫复合物相关疾病的产品、能够判断受试者患所述IgA免疫复合物相关疾病风险的产品、能够诊断所述IgA免疫复合物相关疾病患者在治愈后是否复发的产品。
本发明提出的诊断产品，不仅能诊断患者是否患有IgA免疫复合物相关疾病，还能判断患者的预后，这种诊断产品集多种诊断功能于一体，在临床上具有广泛的应用前景。
附图说明
下面结合附图和实施例对本发明做进一步的说明，其中：
图1为本发明实施例1中提出的试剂盒的检测方法的示意图；
图2为本发明实施例2中对部分儿童医院病例、健康儿童和健康成人的外周血血清中IgA免疫活动性指数检测的结果图；
图3为本发明实施例3对三组IgA肾病患者分别用三种方法治疗后与未治疗组的IgA免疫活动性指数的检测结果对比图；其中，左为第一组患者，采用口服激素治疗；中为第二组患者，采用激素冲击治疗；右为第三组患者，采用免疫抑制剂环磷酰胺治疗；
图4为本发明实施例3中对IgA肾病患者在肾穿刺时、治疗后第一次回访和治疗后第二次回访时的IgA免疫活动性指标、血清中总IgA和缺糖性IgA进行检测的结果图；其中，每根线代表一位患者的不同时间点的检测指标，左为对IgA免疫活动性指数的检测，中为对血清中总IgA的检测，右为对缺糖性IgA的检测；
图5为本发明实施例4中通过检测血清IgA免疫活动性指数来评价对IgA肾病的诊断效果图，具体绘制了受试者操作曲线图；
图6为本发明实施例5中对儿童病例和健康人的外周血血清中IgA免疫活动性指数检测的结果图；
图7为本发明实施例5中通过检测血清IgA免疫活动性指数来评价对过敏性紫癜的诊断效果图，具体绘制了受试者操作曲线图；
图8为本发明实施例5中通过检测血清总IgA来评价对过敏性紫癜的诊断效果图，具体绘制了受试者操作曲线图。
具体实施方式
下面详细描述本发明的实施例，所描述的实施例是示例性的，仅用于解释本发明，而不应理解为对本发明的限制。
本发明的描述中，除非另有明确的限定，提取、包被等词语应做广义理解，所属技术领域技术人员可以结合技术方案的具体内容合理确定上述词语在本发明中的具体含义。
本发明的描述中，参考术语“一个实施例”、“一些实施例”等的描述意指结合该实施例描述的具体特征或材料包含于本发明的至少一个实施例中。在本说明书中，对上述术语的示意性表述不一定指的是相同的实施例。而且，描述的具体特征或材料可以在任何的一个或多个实施例中以合适的方式结合。
实施例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，均可从商业途径得到的试剂和材料。
实施例1
本实施例提供了一种检测IgA免疫复合物(通过检测IgA免疫复合物的浓度来表征IgA免疫活动性指数，IgA免疫活动性指数是指血清/血浆中的IgA免疫复合物的浓度)的试剂盒，该试剂盒中包括分子探针，命名为HQP001，分子探针HQP001为IgA的Fc受体蛋白(FCAR或CD89)。
上述试剂盒包括：分子探针HQP001冻干粉、包被缓冲液、酶标板、标准品、酶标记的二抗、显色液、终止液、洗涤液、稀释液和封闭液。
其中，分子探针HQP001冻干粉为重组FCAR蛋白冻干粉；
包被缓冲液为0.05M的碳酸盐缓冲液(pH＝9.6)；
酶标板选取96孔板；
标准品为商品化的人体血清IgA；
酶标记的二抗为辣根过氧化物酶(HRP)标记的小鼠抗人的IgA；
显色液包括显色液A—3,3’,5,5’-四甲基联苯胺(TMB)和显色液B—过氧化氢溶液；
终止液为10％的硫酸；
洗涤液为含0.05％吐温-20的浓度为0.15M的磷酸盐缓冲液；
稀释液为洗涤液+1‰BSA；
封闭液为洗涤液+5％BSA。
上述试剂盒的检测方法如下，具体流程参加图1：
(1)将分子探针HQP001冻干粉溶解于包被缓冲液中，得分子探针溶液；
(2)将分子探针溶液加入96孔酶标板的每个孔中(图1中表示的是加入96孔板的一个孔中的示意图)，覆盖封口膜，置于4℃环境中过夜；甩去孔中的溶液，用洗涤液洗酶标板多次，向酶标板中加入封闭液，覆盖封口膜，置于4℃环境中过夜，用洗涤液洗酶标板1次，干燥得包被有分子探针HQP001的酶标板；
(3)用稀释液稀释待检测样本得样本稀释液，标准品、阳性对照和阴性对照不用稀释液稀释；然后将样本稀释液、标准品梯度稀释液、阴性对照液、阳性对照液分别加入包被有分 子探针HQP001的酶标板中，覆盖封口膜进行孵育，孵育结束后用洗涤液洗酶标板3次；将HRP标记的二抗加入酶标板中孵育，孵育结束后洗涤酶标板3次；将显色液A和显色液B按1：1的比例混合加入酶标板中，室温避光显色，显色结束后加入终止液，轻轻震荡后将酶标板置于酶标仪中检测吸光值(OD值)，第一波长450nm，第二波长630nm。
实施例2
本实施例使用实施例1提出的试剂盒对肾穿刺确诊患者(IgA肾病)的外周血血清中的IgA免疫活动性指数进行了检测，以验证肾穿刺诊断结果。
图2为对12个儿童医院的IgA肾病患者(通过肾穿刺确诊)、19个健康儿童和13个健康成人的血清中的IgA免疫活动性指数进行了检测。采集上述三类人群的外周血，提取血清，然后用实施例1提出的试剂盒，按实施例1提出的检测方法进行检测，结果如图2所示。图2中，位于10和20之间的水平线为健康人(包括儿童及成年人)的IgA免疫活动性指数的最大值(＝16)，将其设置为门槛线，IgA免疫活动性指数高于门槛线16的为阳性，低于门槛线16的为阴性，其中阳性质控品为浓度为250ng/mL的商品化的人源血清IgA，其中IgA免疫复合物约占1～2％，阴性质控品为水及反应液。结果显示，与标准品、阳性质控品和阴性质控品作为对照，在健康儿童和健康成人中，所有样本的IgA免疫活动性指数均低于门槛线，为阴性；而在儿童医院病例中，其中有4例为阳性，8例为阴性，表明实施例1提供的检测试剂盒和检测方法在真实病例和健康人的检测中是实用的，且准确度高。需要提出的是，随着健康人样本数的增加，取最大值为门槛线可能不稳定，可以用95或者90百分位数(Percentile)取代。另外，对于IgA免疫活动性指数的阴性质控品和阳性质控品可通过如下方式改进：大批量检测健康人群的IgA免疫活动性指数建立正常基础范围：0～平均值+标准差；弱活动性(即IgA免疫活动性指数较低)：平均值+标准差～平均值+2*标准差；强活动性(即IgA免疫活动性指数较高)：>平均值+2*标准差等。
实施例3
本实施例使用实施例1提出的试剂盒对肾穿刺IgA肾病患者用药效果进行了评估。
目前IgA肾病还没有完全治愈的治疗方案，治疗的主要目标为病情控制。病情控制的临床表现体现在三个方面：(一)蛋白尿减少或者消失；(二)肌酐保持长期的稳定，血肌酐正常范围44～133，其中男性53～106，女性44～97；(三)血压正常。对于治疗有效的病人， 治疗过程一般需要六个月至两年才能够控制病情，亦即上述三个指标改善并到达正常范围。但IgA肾病患者中40％最终发展为肾衰竭(End Stage Kidney Disease，ESKD)。由此可见，上述临床指标对治疗效果的反应时间滞后3-6个月。
而IgA免疫活动性指数直接反映治疗过程中IgA免疫的抑制效果，没有时间滞后性，可以作为治疗效果的互补指标；另一方面，基于外周血血清的IgA免疫活动性指数操作简单，检测费用低廉，通过多个时间段的抽血检测，建立病人IgA免疫活动性随治疗流程的时间变化，可以帮助医生实时跟踪病人体内IgA免疫活动性的起起伏伏，把握病情控制的即时证据及治疗效果的动态走势。
对于可能为IgA肾病的病人，可在肾穿刺前采集外周血，提取血清，按照实施例1中IgA免疫活动性指数试剂盒操作流程，建立病例的IgA免疫活动性指数基线。在肾穿刺确诊后，按照时间间隔分别采集外周血进行IgA免疫活动性指数检测，比如治疗开始的第一天，第二天，第三天，第七天，第三十天，第六十天，第九十天，以此类推，分别采集外周血进行IgA免疫活动性指数检测；对于需要二次或二次以上疗程的，在每个新疗程开始后的第一天，第二天，第三天，第七天，第三十天，第六十天，第九十天，以此类推，分别采集外周血进行IgA免疫活动性指数检测。经济条件宽裕的可以酌情增加检测频率，经济条件受限的可以酌情减少检测频率；治疗结束后应该每隔一个月，或者每隔三个月进行外周血采集来检测IgA免疫活动性指数。由此描绘IgA免疫活动性指数的时间动态曲线。
本实施例进行了一个早期临床研究，收集穿刺确诊的IgA肾病患者174例，根据患者的身体指标，将上述患者分为四组，其中一组80例没有治疗，对其他三组患者进行不同的治疗，第一组45例患者采用口服激素的方法进行治疗，第二组17例患者采用激素冲击进行治疗，第三组32例患者采用免疫抑制剂环磷酰胺进行治疗。在穿刺确诊时对三组患者的外周血进行一次采集，提取血清，用实施例1中的试剂盒及检测方法分别对三组患者的血清IgA免疫活动性指数进行检测，记录检测值；对三组患者进行相应的治疗后，每隔至少3个月回访一次，回访时采集三组患者的外周血，进行一次IgA免疫活动性指数的检测，同时对未治疗组患者的血清IgA免疫活动性指数也进行检测，记录检测值；对未治疗组和治疗后的3组患者的IgA免疫活动性指数检测值进行统计，结果如图3所示。用三个治疗组的第一次回访与未治疗组分别对比，图3中，左为第一组患者治疗后的情况，采用口服激素治疗；中为第二组患者治 疗后的情况，采用激素冲击治疗；右为第三组患者治疗后的情况，采用免疫抑制剂环磷酰胺治疗。从图3可以看出，三种方案治疗至少3个月后的外周血IgA免疫活动性指数，其人群平均值比未治疗组均下降超过50％，且有统计学意义，表明三种治疗方案对于控制IgA免疫反应均具有显著效果。
本实施例还从上述患者中随机选出37位患者，对每一位患者进行贯序跟踪，对三个时间点，分别为肾穿刺时、治疗后第一次回访和治疗后第二次回访，采用实施例1中的试剂盒及检测方法对IgA免疫活动性指数进行检测作为基准线，采用人血清/血浆总IgA试剂盒(ELISA)及半乳糖缺乏型IgA1(Gd-IgA1，为IgA肾病的标志物)特异性抗体(KM55)检测试剂盒分别对血清中总IgA和缺糖性IgA(Gd-IgA)进行检测，结果如图4所示。图4中，每根线代表一位患者的不同时间点的检测指标，其中左为对IgA免疫活动性指数的检测，中为对血清中总IgA的检测，右为对缺糖性IgA的检测。图4左表明IgA免疫活动性指数在治疗过程中持续下降，尽管第一次回访与基准线对比具有统计重要性，表明治疗过程中病情在改善；而总体IgA及缺糖性IgA并没有显示病情改善，总体IgA也体现了下降趋势，但第一次回访及第二次回访分别与基准线对比统计学意义不明显；缺糖性IgA反而出现上升趋势，这可能是因为缺糖性IgA检测技术的不可靠性。
值得一提的是，在同一个临床试验中，IgA免疫活动性指数还体现出与尿常规异常指标及临床病理检测的关联性。免疫活动性指数高的人群比指数低的人群肾小球滤过率(eGFR)平均下降10％；对于肾穿刺后病理诊断的牛津分型，IgA免疫活动性指数高的人群毛细血管内增生(E1)比例或肾小管萎缩/间质纤维化的严重程度(T1/2)比例均超过指数低人群两倍以上，表明IgA肾病患者的IgA免疫活动性指数越高，肾损伤越严重。
实施例4
本实施例使用实施例1提出的试剂盒评估了血常规、尿常规或肾功能指标异常的病人患IgA肾病的风险。
IgA肾病为IgA免疫复合物在肾脏不同组织沉积导致局部免疫反应的肾炎，可能涉及肾小球，肾小管间质及肾血管，且80％的人为青壮年(20-35岁)，在IgA肾病发病初期，微弱的血常规或尿常规指标异常或肾功能指标异常容易被医生与病人忽视，对于未来通过肾穿刺确诊的病人，其肾损伤从开始可以持续3～5年才表现出明显症状。
本实施例利用实施例1提出的试剂盒及检测方法对比研究了108例IgA肾病病人(IgAN)与63个健康人(Healthy Control，HC)的血清IgA免疫活动性指数，t-检测显示两个人群的IgA免疫活动性指数具有显著差异，对应的受试者操作曲线(ROC)图见图5，曲线下面积(AUC)为0.89，最优的门槛线对应特异性为75％，敏感性为89％，表明通过检测血清IgA免疫活动性指数来诊断患有IgA肾病的风险是有参考价值的。
可见，使用IgA免疫活动性指数可较为准确地评估血常规或尿常规或肾功能指标异常的病人患IgA肾病的风险，IgA免疫活动性指数偏高可以立刻引起医生与病人的重视，预警IgA肾病的高风险，从而让病人开始进行健康管理，尽量延缓肾损伤。
实施例5
本实施例使用实施例1提出的试剂盒病人是否患有过敏性紫癜进行了诊断。
目前临床上对过敏性紫癜诊断的几大要点为：(一)血常规检查：全血白细胞及嗜酸性粒细胞增高，出血严重时，红细胞及血红蛋白降低；(二)典型特征性皮肤紫癜，结合关节、胃肠或肾脏症状以及反复发作史；(三)多有感染、食物、药物、花粉、虫咬、疫苗接种等病史；(四)血沉增快，血清C-反应蛋白(CPR)可呈阳性，血清IgA增高；(五)伴随肾损伤的有尿蛋白及尿血。对过敏性紫癜快速确诊可以减缓可能的肾损伤。(四)中血清IgA增高为过敏性紫癜的特征，但IgA增高对紫癜特异性不高，因为许多其它感染性疾病也可以导致IgA上升。由于过敏性紫癜为IgA复合物在血管导致的炎症损伤，血清IgA免疫活动性指数显示出对过敏性紫癜较高的特异性。
本实施例采用实施例1中的试剂盒对比研究了157例过敏性紫癜儿童(HSPN)与33个健康人(HC，包括儿童及成年人)的血清IgA免疫活动性指数，HSPN(n＝157)与HC(n＝33)人群的IgA免疫活动性指数平均值分别为18.82，9.15，HSPN几乎为HC的两倍，且t-检验的p-值为2E-23，差异的统计学意义强烈。通过采集上述过敏性紫癜和健康人的外周血，提取血清，然后用实施例1提出的试剂及检测方法对IgA免疫活动性指数进行检测，结果如图6所示。图6中，位于10和20之间的水平线为健康人的IgA免疫活动性指数的最大值，将其设置为门槛线，IgA免疫活动性指数高于门槛线的为阳性样本，阳性质控品为浓度为250ng/mL的商品化的人源血清IgA(其中IgA免疫复合物占1～2％)，阴性质控品为水及反应液。可见大部分过敏性紫癜儿童的IgA免疫活动性指数超过正常人，且大部分为正常人的2 倍或2倍以上。
对比试验的受试者操作曲线(ROC)图如图7所示，曲线下面积(AUC)为0.94，最佳门槛值对应的特异性为85％，敏感性为85％，表明通过检测血清IgA免疫活动性指数来诊断是否患有过敏性紫癜是有参考价值的。
而对传统的血清总IgA统计分析发现，152例过敏性紫癜儿童(HSPN)与8个健康儿童(HC)中，HSPN(n＝152)与HC(n＝8)人群的总IgA信号平均值分别为4.81，2.99，HSPN只高出HC大约61％，t-检验的p-值为0.0035，也有统计重要性。对应的受试者操作曲线(ROC)图如图8所示，曲线下面积(AUC)为0.71，最佳门槛值对应的特异性为75％，敏感性为72％。
上述血清IgA免疫活动性指数与总IgA用于诊断过敏性紫癜的对比结果显示，IgA免疫活动性指数作为过敏性紫癜诊断生物标志物，其临床效果远远优于总IgA(具体数值见表1)。
表1
Figure PCTCN2022127450-appb-000001
综上，在临床诊断的第(四)点中，用检测血清IgA免疫活动性指数取代总IgA，可以帮助医生对满足其它诊断要点、且血清IgA免疫活动性指数增高的病人快速确诊。
上面对本发明实施例作了详细说明，但是本发明不限于上述实施例，在所属技术领域普通技术人员所具备的知识范围内，还可以在不脱离本发明宗旨的前提下作出各种变化。此外，在不冲突的情况下，本发明的实施例及实施例中的特征可以相互组合。

Claims (10)

  1. 检测IgA免疫复合物的试剂在制备IgA肾病诊断产品中的应用，其特征在于，所述试剂包括分子探针，所述分子探针包括IgA的Fc受体蛋白，用于特异性结合所述IgA免疫复合物。
  2. 根据权利要求1所述的应用，其特征在于，所述分子探针为溶液或冻干粉。
  3. 根据权利要求2所述的应用，其特征在于，所述产品包括试剂盒、芯片或试纸中的任一种。
  4. 根据权利要求3所述的应用，其特征在于，所述试剂盒包括分子探针冻干粉、包被缓冲液、酶标板；所述包被缓冲液通过溶解所述分子探针冻干粉以将所述分子探针包被于所述酶标板上，用于特异性结合所述IgA免疫复合物。
  5. 检测IgA免疫复合物的试剂在制备过敏性紫癜诊断产品中的应用，其特征在于，所述试剂包括分子探针，所述分子探针包括IgA的Fc受体蛋白，用于特异性结合所述IgA免疫复合物。
  6. 根据权利要求5所述的应用，其特征在于，所述分子探针为溶液或冻干粉。
  7. 根据权利要求5所述的应用，其特征在于，所述产品包括试剂盒、芯片或试纸中的任一种。
  8. 一种IgA免疫复合物相关疾病的诊断产品，其特征在于，所述产品包括检测IgA免疫复合物的试剂，所述试剂包括分子探针，所述分子探针包括IgA的Fc受体蛋白。
  9. 根据权利要求8所述的诊断产品，其特征在于，所述产品包括试剂盒、芯片或试纸中的任一种。
  10. 根据权利要求8所述的诊断产品，其特征在于，所述产品包括能够判断受试者是否患有所述IgA免疫复合物相关疾病的产品、能够判断受试者患所述IgA免疫复合物相关疾病风险的产品、能够诊断所述IgA免疫复合物相关疾病患者在治愈后是否复发的产品。
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