妊娠相关血清蛋白A光激化学发光免疫分析法的建立  

Development of an amplified luminescent proximity homogeneous immunoassay kit for the detection of pregnancy-associated plasma protein-A in human serum

作　　者：黄莉萍[1] 周燕莉[1] 邹丽萍[2] 刘天才[2] 董志宁[2] 钟梅[1] 吴英松[2] 

机构地区：[1]南方医科大学南方医院妇产科,广东广州510515 [2]南方医科大学抗体工程研究所,广东广州510515

出　　处：《分子诊断与治疗杂志》2014年第1期37-41,共5页Journal of Molecular Diagnostics and Therapy

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（AlphaLISA）建立快速定量检测妊娠相关血清蛋白A（PAPPA）的方法。方法用两株配对的PAPPA单克隆抗体，一株PAPPA单克隆抗体包被受体微球，另一株PAPPA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PAPPA试剂分析灵敏度为1．365mU／L，线性测量范围为1．365～10000mU／L，分析内和分析间的精密度分别为5．61％～6．51％和2．07％～4．82％，与AFP、RPL、HPL和HCG无明显交叉反应。使用自制试剂对853份9～13周的正常孕妇血清进行检测，建立了孕妇血清在不同孕周时的中位数水平，为正常值参考范围的确定提供了一定依据，其中165份临床血清样本使用自制试剂与Perkin Elmer公司PAPPATRFIA诊断试剂盒同时进行检测，结果具有相关性（r=0．987）。结论自制PAPPA光激化学发光免疫分析试剂的各项性能指标均能达到临床检测要求，有望替代国外同类产品。Objective To establish an amplified luminescent proximity homogeneous immunoassay （AlphaLISA） for the determination of pregnancy-associated plasma protein-A （PAPPA） in human serum. Methods Two anti-PAPPA monoclonal antibodies were used to develop the sandwich AlphaLISA kit for detection of PAPPA in serum. One was coated on AlphaLISA acceptor beads and the other was biotinylated. The kit also contains donor beads coated with streptavidin. The optimal test conditions and analytical performance of the method were studied. Results The analytical sensitivity of AlphaLISA for PAPPA was 1.365 mU/L. The measure range of developed kit was 1.365-10 000 mU/L. The intra-assay and inter-assay coefficient of variation were 5.61%-6.51% and 2.07%-4.82%, respectively. There was no cross-reaction to AFP, RPL, HPL and HCG. 853 maternal serum samples of nine to thirteen pregnant weeks were tested using reagents made by ourselves. Median values during different weeks and the reference value were established. The correlation coefficient of blood samples detection results between home-made and commercial PAPPA TRFIA kits （PerkinElmer） was 0.987. Conclusion This new developed PAPPA AlphaLISA kit is a valuable test for clinical application with better sensitivity, specificity, and accuracy.

关 键 词：妊娠相关血清蛋白A 光激化学发光 测定 

分 类 号：R714.1[医药卫生—妇产科学]