人促卵泡激素光激化学发光免疫分析法的建立及性能评价

检验医学 ›› 2012, Vol. 27 ›› Issue (9): 736-740.

人促卵泡激素光激化学发光免疫分析法的建立及性能评价

林冠峰,董志宁,贺安,邹丽萍,侯经远,李明,吴英松

南方医科大学生物技术学院，广东广州 510515

收稿日期:2012-05-17 修回日期:2012-05-08 出版日期:2012-09-12 发布日期:2012-09-12
通讯作者: 吴英松，联系电话：020-61648321。
作者简介:林冠峰，男，1988年生，学士，主要从事新型免疫检测技术的研究。
基金资助:
国家科技重大专项基金资助项目(2009ZX10004-706)

Establishment and performance evaluation of AlphaLISA for the detection of human follicle stimulating hormone

College of Biotechnology，Southern Medical University，Guangdong Guangzhou 501515，China

Received:2012-05-17 Revised:2012-05-08 Online:2012-09-12 Published:2012-09-12

摘要/Abstract

摘要： 目的　利用光激化学发光免疫分析法（AlphaLISA）建立人促卵泡激素(hFSH)的快速检测试剂。方法　使用2株配对的hFSH单克隆抗体，一株hFSH单克隆抗体用生物素标记，另一株hFSH单克隆抗体包被于受体微球上，与包被有链霉亲合素的供体微球共同构成检测试剂，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hFSH试剂分析敏感性和功能敏感性分别为0.09和0.12 U/L，线性测量范围为0.09~192.00 U/L，试剂的分析内变异系数（CV）和分析间CV分别为4.2%~7.1%和7.5%~8.3%，均<10.0%，与人黄体生成素(hLH)、人促甲状腺素(hTSH) 和人绒毛膜促性腺激素(hCG) 的交叉率分别为0.16%、0.08%和0.02%。100份临床血清样本用本试剂与西门子 Immulite 2000 促卵泡生成激素化学发光（CLIA）检测试剂盒检测，其相关系数为 0.979(P<0.001)。结论　自制hFSH AlphaLISA试剂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hFSH浓度的测定。

关键词: 人促卵泡激素, 光激化学发光免疫分析法, 性能评价

Abstract: Objective　To develop rapid amplified luminescent proximity homogeneous immunoassay (AlphaLISA) reagent for the detection of human follicle stimulating hormone(hFSH). Methods　Two monoclonal anti-hFSH antibodies were used to develop the AlphaLISA reagent for the detection of hFSH. One was biotinylated，and the other was coated on AlphaLISA acceptor beads. The reagent also contained donor beads coated with streptavidin. The optimal test conditions and analytical performance of the method were evaluated. Results　The analytical sensitivity and functional sensitivity of AlphaLISA for the detection of hFSH were 0.09 and 0.12 U/L. The linear measurement range of the developed reagent was 0.09-192.00 U/L. The intra- and inter-assay coefficients of variation (CV) were 4.2%-7.1% and 7.5%-8.3%，which were all<10.0%，respectively. The ratios of cross-reactivity were 0.16%，0.08% and 0.02% with human luteinizing hormone(hLH)，human thyroid stimulating hormone(hTSH) and human chorionic gonadotropin(hCG)，respectively. The results of 100 samples′ detection by this reagent showed good correlation coefficient with those of SIEMENS Immulite 2000 follicle stimulating hormone kit by chemiluminescent immunoassay (CLIA，r=0.979，P<0.001). Conclusions　The developed hFSH AlphaLISA reagent in this study meets the requirement of clinical determination，and is a valuable test for clinical application.

林冠峰;董志宁;贺　安;邹丽萍;侯经远;李　明;吴英松. 人促卵泡激素光激化学发光免疫分析法的建立及性能评价[J]. 检验医学, 2012, 27(9): 736-740.

LIN Guanfeng;DONG Zhining;HE An;ZOU Liping;HOU Jingyuan;LI Ming;WU Yingsong.. Establishment and performance evaluation of AlphaLISA for the detection of human follicle stimulating hormone[J]. , 2012, 27(9): 736-740.

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