微板式化学发光酶免疫分析法测定人血清中癌胚抗原  

Microplate Chemiluminescence Enzyme Immunoassay for the Determination of Carcinoembryonic Antigen in Human Serum

作　　者：辛天兵[1] 王栩 靳辉[1] 车景辉 梁淑轩[1] 李振甲[3] 林金明[2] 

机构地区：[1]河北大学化学与环境科学学院,保定071002 [2]清华大学化学系,北京100084 [3]北京科美东雅生物技术有限公司,北京100094

出　　处：《分析化学》2008年第8期1056-1060,共5页Chinese Journal of Analytical Chemistry

基　　金：国家973计划(No.2007CB714507);863计划(No.2006AA02Z4A8)资助项目

摘　　要：采用辣根过氧化物酶(HRP)催化鲁米诺(luminol)-H2O2化学发光体系,建立了一种测定人血清中癌胚抗原(CEA)的高灵敏度、高特异性、简便快速的微板式化学发光酶免疫分析方法。对免疫反应条件、酶结合物稀释度、发光反应时间、封闭液等进行了考察和优化。采用双抗体夹心法,室温静置1h,洗涤后加入100μL发光底物液,10min后检测。该方法的线性相关系数为0.9998;最低检出限为0.57μg/L;批内和批间变异均在10%之内;低、中、高3个不同浓度值样品的平均回收率分别为107.4%、93.3%和104.5%。使用本方法与进口发光试剂盒对40份人血清样品进行测定,结果表明,本方法显示了良好的相关性,其相关系数为0.9115。表明本分析体系稳定可靠,可用于商品化诊断试剂盒的开发和应用。A highly sensitive, specific, simple and rapid microplate chemiluminescence enzyme immunoassay （CLEIA） was developed for the determination of carcinoembryonic antigen （CEA） in human serum, which is based on the luminol-H2O2 reaction catalyzed by horseradish peroxidase （HRP） as the chemiluminescence system. Several physicochemical parameters such as the immunoreaction conditions, the dilution ratio of CEA- HRP, chemiluminescent time, and blocking solutions were studied and optimized. A sandwich reaction was performed with incubation at room temperature for 1 h, followed by adding 100 μL chemiluminescent substrate and measuring after 10 min incubation. The correlation coefficient of the proposed method for CEA was 0. 9998. The detection limit was 0.57 μg/L. The RSD of inter-assay and intra-assay were both less than 10%. The average recoveries of three different spiked concentration samples were 93.3% - 107.4%. The proposed method has been successfully applied to the determination of CEA in 40 human sera and has shown a good correlation compared with the commercially CLIA kit with a correlation coefficient of 0. 9115. This method has shown great potential in the fabrication of diagnostic kit and could be used in the clinical analysis of CEA in human serum.

关 键 词：化学发光酶免疫分析 辣根过氧化物酶 癌胚抗原 人血清 

分 类 号：R446.6[医药卫生—诊断学]