β_2-微球蛋白酶联免疫分析试剂盒的研制

Establishment of an Enzyme Linked Immunosorbent Assay for β_2-Microglobin

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0.15mg/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=0.997 4。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5mg/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yELIA=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。

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 horse- radish peroxidase（HRP）. TMB-H2 02 solution was used as the substrate of HRP. The sensi- tivity of the assay is 0. 15 mg/L. The intra- and inter-assay CVs were 4.06%-11.2% and 14.1%-16.0%, respectively. The analytical recovery was 93.3%-115.9%, the coefficient correlation was 0. 997 4. The assay is specific for β2-MG and no cross reaction with Fer, lit- tle cross reaction with albumin and u-fetoprotein when it is below 10 mg/L and 5 mg/L, re- spectively. For a reference RIA method, the correlation regression equation of present method is yELISA =1-006ZVRIA＋0. 406, r=0. 900（n=59）. This method for measuring β2-MG is rapid, sensitive and convenient. It is suitable for clinical and research application.