人附睾分泌蛋白4光激化学发光免疫分析法的建立  

Development of Alpha LISA for the detection of luteinizing hormone in human serum

作　　者：肖俐霞 林冠峰[1] 陈少琅[1] 吴英松[1] 

机构地区：[1]南方医科大学生物技术学院,广东广州510515

出　　处：《热带医学杂志》2015年第5期592-595,共4页Journal of Tropical Medicine

基　　金：国家科技重大专项(2009ZX10004-706)

摘　　要：目的采用光激化学发光免疫分析技术建立快速定量检测人附睾分泌蛋白4（HE4）的方法。方法用两株配对的HE4单克隆抗体,一株HE4单克隆抗体包被受体微球,另一株HE4单克隆抗体用生物素标记,与链霉亲合素的供体微球共同组成检测试剂。结果自制HE4试剂分析灵敏度为0.81 pmol/L,分析内和分析间的精密度分别为4.3%~7.2%和6.1%~7.7%,均低于10%,与癌抗原15-3（CA15-3）、甲胎蛋白（AFP）、癌胚抗原（CEA）、癌抗原125（CA125）无明显交叉反应,120份临床血清样本用本试剂与罗氏电化学发光检测试剂盒平行检测,其相关系数为0.95。结论自制HE4光激化学发光免疫分析试剂各项指标均能达到临床要求,有望替代国外同类产品。Objective To establish an amplified luminescent proximity homogeneous immunoassay（Alpha） for determination of human epididymis protein 4（HE4） in human serum. Methods Two anti-HE4 monoclonal antibodies were used to develop Alpha LISA for the detection of human epididymis protein 4 in human serum. One was coated on Alpha LISA acceptor magnetic beads and the other was biotinylated. The reagent also contained magnetic beads coated with streptavidin. The optimal test conditions and analytical performance of the method were studied. Results The analytical sensitivity of Alpha LISA for HE4 was 0.81 pmol / L. The intra- and inter-assay coefficient of variation were all less than 10%. There was no cross-reaction to cancer antigen 15-3（CA15-3）,alpha fetoprotein（AFP）,carcino-embryonic antigen（ CEA）,cancer antigen 125（CA125）. Results of 120 samples detection by this reagent showed good correlation coefficient with ECLIA HE4 kit（Roche）. Conclusion The developed HE4 Alpha LISA reagent in this study was a valuable test for clinical application with better sensitivity, specificity, and accuracy.

关 键 词：人附睾分泌蛋白4 光激化学发光 免疫分析 

分 类 号：R392-33[医药卫生—免疫学]