β2-微球蛋白酶联免疫分析试剂盒的研制

摘要

A sensitive and specific ELISA for β2-Microglobin was established by using anti-β2-Microglobin antibody coated on the microtiter plate, and β2-Microglobin labeled horseradish peroxidase(HRP). TMB-H2O2 solution was used as the substrate of HRP. The sensitivity of the assay is 0. 15 mg/L. The intra- and inter-assay CVs were 4. 06%-ll, 2% and 14. l%-16. 0%, -respectively. The analytical recovery was 93. 3%-115. 9%, the coefficient correlation was 0. 997 4. The assay is specific for JB2-MG and no cross reaction with Fer, little cross reaction.with albumin and a-fetoprotein when it is below 10 mg/L and 5 mg/L, respectively. For a reference RIA method, the correlation regression equation of present method is YELIsa = 1. 006xR1A+0. 406, r = G. 900(n = 59). This method for measuring β2-MG is rapid, sensitive and convenient. It is suitable for clinical and research application.%以β2-微球蛋白(β2-MG)抗体包被微孔板,四甲基联苯胺(TMB)为底物,用辣根过氧化物酶标记β2-MG,建立了β2-微球蛋白酶联免疫分析试剂盒,并对该方法进行了方法学分析.结果显示:本方法分析灵敏度为0.15 mg/L,批内,批间变异系数分别为4.1％～11.2％和14.1％～16.0％,回收率为93.3％～115.9％,高浓度β2-MG样品系列倍比稀释后,测定值与稀释度呈线性相关,相关系数为r=0.9974.特异性结果显示:本试剂盒与铁蛋白基本无交叉反应,与白蛋白及甲胎蛋白在浓度分别低于10mg/L及5 mg/L时有交叉反应,但反应很小.与放射免疫分析法(RIA)同时测定人血清样品,方法间有良好的相关性,相关方程为yε1.1A=1.006xRIA+0.406,r=0.900(n=59).以上参数均符合临床免疫分析的要求.本试剂盒操作简便、快速,适用于临床检测和科研应用.