【摘要】 目的:建立一种检测人血清中免疫球蛋白A1(IgA1)的酶联免疫吸附方法,用于血清学法诊断IgA肾病(IgAN)病程进展。方法:以特异性抗人IgA1单克隆抗体(M cAb)为包被抗体,辣根过氧化物酶(HRP)标记的抗人IgA作为标记抗体建立人血清IgA1ELISA法,并对该技术进行一系列反应条件优化实验,用于临床初步检测IgAN患者血清IgA1的水平。结果:实验优化的包被抗体浓度为1.0μg/m l,血清稀释倍数1:1000,HRP标记抗体的最佳工作浓度为1:4000。本方法特异性强,灵敏度高,精密度好。该法测定范围10~500 mg/dL,最低检出量10 mg/dL,批内和批间CV%分别为6.8%和9.3%。健康对照组测定值:139.0±55.2 mg/d l(n=150),IgAN确诊病人:207.4±59.3(n=163),两者差异有显著意义(P<0.01);IgAN疑诊病人组的测定值:168.6±47.4(n=185),高于正常对照组。结论:血清IgA1-ELISA检测方法,具有灵敏度高、特异性好的优点,可以作为IgAN诊断的辅助手段。更多还原

【Abstract】 Objective:To establish an enzyme-linked immunosorbent assay(ELISA) for immunoglobin A1(IgA1) in human serum,applied to the serological diagnosis of the progression of IgAN.Methods:A Detecting system was established,including anti-IgA1 McAb(Coating antibody) and polyclone anti-IgA antibody labeled with HRP(detecting antibody).After a series of optimized tests have completed,the kit was used to detect serum level of IgA1 in IgAN patients.Results:Experiment showed that the optimal concentration of the coating antibody,diluted ratio of serum and detecting antibody with HRP were 1 μg/ml,1:1000 and 1:4000,respectively.This assay had good reproducibility and steadiness,detective range was from 10 mg/dl to 500 mg/dl,the sensitivity reached 10 mg/dl.Within day and between day coefficients of variation were 6.8% and 9.3%,respectively.The results values from 150 controls,163 patients with IgAN and 185 patients with uncertain IgAN were 139.0±55.2 mg/dl,207.4±59.3 and 168.6±47.4,respectively.The significance between the controls and the patients with IgAN was observed(P<0.01).Conclusion:This assay is sensitive,specific and valuable for the diagnosis of patients with IgA nephropathy.更多还原