血清人附睾分泌蛋白-4(HE4)时间分辨免疫荧光分析检测试剂的研制  

Development of time-resolved fluoroimmunoassay kit for detection of human epididymis protein 4(HE4) in serum

作　　者：陈少琅[1] 万勇[1] 张琼[1] 周健伟[1] 徐伟文[1] 吴英松[1] 

机构地区：[1]南方医科大学生物技术学院抗体工程研究所,广东广州510515

出　　处：《热带医学杂志》2014年第4期446-449,520,共5页Journal of Tropical Medicine

基　　金：国家863计划(SS2012AA020506)

摘　　要：目的利用时间分辨免疫荧光分析技术（TRFIA）建立血清人附睾分泌蛋白-4（HE4）的检测试剂。方法采用双抗体夹心法建立血清HE4-TRFIA检测试剂并对试剂的各项指标进行评价。结果HE4-TRFIA检测试剂的灵敏度为0．73pmol／L，测量范围为0．73～1000pmol／L，分析内和分析间的精密度分别为4．6％～7．5％和8．3％～14．4％。与CA15．3、CEA、AFP、HBsAg和CA125无交叉反应。317份正常女性血清标本测试结果提示本试剂的正常参考值范围是0～90pmol／L。与进口的同类试剂检测，检测结果的相关系数R^2为0．976。结论本HE4-TRFIA检测试剂各项指标均能达到临床检测要求，可望替代国外同类产品用于临床检测。Objective To establish a kit for detection of human epididymis protein 4 （HE4） in serum by time-resolved fluoroimmunoassay （TRFIA）. Method Time-resolved fluoroimmunoassay kit for detection of HE4 in serum was developed based on the double-antibody sandwich method. Results The analytical sensitivity of the kit was 0.73 pmol/L.The measure range of developed kit was 0.73-1 000 pmol/L. The intra- and inter-assay coefficients of variation （CV） were 4.6%-7.5% and 8.3%-14.4%, respectively. There was no cross reactivity with cancer antigenlS-3（CA15-3）,carcinoembryonic antigen（CEA）, alpha-fetoprotein （AFP）, hepatitis B virus surface antigen （HBsAg） and cancer antigen 125（CA125）. The cut-off value for TRFIA HE4 kit in healthy female subjects （n=317） was 90 pmol/L. The correlation coefficient of blood samples detection results between this kit and a commercial electrochemiluminescence immunoassay （ECLIA） kits was 0.976. Conclusion This newly developed HE4-TRFIA kit is valuable for clinical application with better sensitivity, specificity, and accuracy.

关 键 词：人附睾分泌蛋白-4 时间分辨免疫荧光分析(TRFIA) 检测 

分 类 号：R737.31[医药卫生—肿瘤]