AFP/CEA双标记时间分辨荧光免疫分析试剂的研制及性能鉴定  

Preparation and identification of dual-label time-resolved fluoroimmunoassay for simultaneous detection of alpha fetoprotein and carcinoembryonic antigen

作　　者：王征[1] 李来庆[1] 杨凤爱[1] 李明[1] 吴英松[1] 

机构地区：[1]南方医科大学生物技术学院,广州510515

出　　处：《山东医药》2010年第34期17-19,共3页Shandong Medical Journal

基　　金：国家863计划资助项目(2006AA02A311)

摘　　要：目的研制甲胎蛋白（AFP）及癌胚抗原（CEA）的双标记时间分辨荧光免疫分析（TRFIA）试剂,并鉴定其性能。方法将抗AFP单克隆抗体（E014）与抗CEA单克隆抗体（3E6）混合包被96孔板,用钐标记抗AFP单克隆抗体（E010）,铕标记抗CEA单克隆抗体（S001）,用双抗体夹心一步法建立AFP/CEA TRFIA试剂,测定其线性相关性、灵敏度、精密度、回收率、特异性,并与对应的单标记进口试剂进行比较。结果 AFP分析灵敏度为0.3 U/ml,线性范围为0.3~800 U/ml,平均回收率为99.8%,分析内和分析间变异系数分别为2.1%~5.8%、4.1%~8.7%;CEA分别为0.2 ng/ml、0.2~400 ng/ml、100.3%、2.4%~5.6%、4.8%~9.8%。该试剂与对应的单标记进口试剂的相关系数分别为0.99、0.98。结论成功研制了AFP/CEA双标记TRFIA试剂,其性能均达到临床检测要求,可替代国内外的单标记试剂。Objective To establish and identify dual-label time-resolved fluoroimmunoassay（TRFIA） for simultaneously detecting alpha fetoprotein（AFP）and carcinoembryonic antigen（CEA） in serum.Methods The mixture monoclonal antibody（McAb） of anti-AFP（E014） and anti-CEA（3E6） were co-coated in the 96 microtitration wells.The anti-AFP McAb（E010） was labeled with Sm3＋-chelates and anti-CEA McAb（S001） was labeled with Eu3＋-chelates.The dual-label time-resolved fluoroimmunoassay were developed by one-step double-antibody sandwich.Results The assay sensitivity for AFP was 0.3 U/ml and the measurable ranges of was 0.3～800 U/ml,and the recovery rate was 99.8%.The intra-and inter-assay coefficient of variation（CV） were 2.1%～5.8% and 4.1%～8.7%,respectively.The assay sensitivity for CEA was 0.2 U/ml and the measurable ranges of was 0.2～400 U/ml.The recovery rate was 100.3%.The intra-and inter-assay CV were 2.4%～5.6% and 4.8%～9.8%,respectively.The correlation coefficient of 100 blood samples detection results between home-made dual-label TRFIA and commercially available single-label DELFIA kit（PerKin Elmer） were 0.99 for AFP and 0.98 for CEA,respectively.Conclusions The home-made AFP/CEA TRFIA kit is valuable for clinical application with very satisfactory sensitivity,specificity,and accuracy.It may be used to replace the single-label reagents for diagnosis of primary hepatic carcinoma or large-scale general survey and screen of liver disease.

关 键 词：免疫酶技术 时间分辨荧光免疫分析技术 甲胎蛋白 癌胚抗原 

分 类 号：R608[医药卫生—外科学]