人促黄体生成素光激化学发光免疫分析法的建立

Development of AlphaLISA for the detection of luteinizing hormone in human serum

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促黄体生成素(hLH)的方法。方法用2株配对的hLH单克隆抗体,1株hLH单克隆抗体包被受体微球,另1株hLH单克隆抗体先用生物素标记,再与链霉亲合素的供体微球共同组成人促黄体生成素光激化学发光免疫分析试剂,进而优化反应体系并对试剂的各项性能指标进行评价。结果自制hLH试剂分析灵敏度为0.164 U/L,线性测量范围为0.164～135 U/L,分析内和分析间的精密度分别为4.3%～6.1%和7.4%～8.1%,均低于10%,与hTSH、hFSH和hCG无明显交叉反应,158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,对所测数值采用配对t检验分析,结果显示两种方法检测结果差异无统计学意义(t=-1.468,P=0.07)。结论自制hLH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 luteinizing hormone（hLH） in human serum.Methods Two anti-hLH monoclonal antibodies were used to develop AlphaLISA for the detection of luteinizing hormone in human serum.One was coated on AlphaLISA acceptor beads and the other was biotinylated.The reagent 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 hLH was 0.164 U/L.The detection range of developed reagent was 0.164～135 U/L.The intra-and inter-assay coefficient of variation was all less than 10%.There was no cross-reaction to hTSH,hFSH and hCG.The result of paired-samples t test of the measured values of 158 clinical serum samples was t=-1.468（P=0.07）,and indicated that there was no significant difference between the detection of hLH-AlphaLISA reagent and commercial ECLIA hLH kit（Roche）.Conclusion The developed hLH AlphaLISA reagent in this study was a valuable test for clinical application with better sensitivity,specificity,and accuracy.