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研制检测人血清神经元特异性烯醇化酶(NSE)的光激化学发光免疫分析(AlphaLISA)快速检测试剂盒。采用受体微球用一株NSE单克隆抗体包被,用生物素标记一株单克隆抗体,与链霉亲和素的供体微球共同组成检测试剂盒,优化反应体系并对试剂盒的各项性能指标进行评价。结果显示:自制NSE试剂盒灵敏度为0.149 ng/ml,线性范围为0.149~600ng/ml,分析内、分析间的精密度分别为3.7%~4.3%、3.9%~5.2%,与细胞角蛋白19片段(CYFRA21-1)、非神经元性烯醇化酶(NNE)均无交叉反应,154份临床血清样本用本试剂盒与罗氏化学发光试剂盒检测,其相关系数为0.979。发现自制NSE-AlphaLISA试剂盒的各项性能指标均能达到临床检测要求,可用于临床血清样本NSE浓度的测定。
Development of a rapid chemiluminescent chemiluminescence immunoassay (AlphaLISA) kit for the detection of human serum neuron-specific enolase (NSE). Recipient microspheres were coated with one NSE monoclonal antibody, one monoclonal antibody was labeled with biotin, the detection kit was composed with the donor microspheres of streptavidin, the reaction system was optimized and the Various performance indicators for evaluation. The results showed that the sensitivity of the NSE kit was 0.149 ng / ml and the linear range was 0.149-600 ng / ml. The precision of the assay was 3.7% -4.3%, 3.9% -5.2% There was no cross-reaction between the fragment (CYFRA21-1) and non-neuronal enolase (NNE), and 154 clinical serum samples were detected by Roche Chemiluminescence Kit with a correlation coefficient of 0.979. Found that the performance of self-made NSE-AlphaLISA kit can meet the clinical testing requirements, can be used for clinical serum samples NSE concentration determination.