微板式化学发光酶免疫分析法测定人血清中癌胚抗原 | |
辛天兵 ; 王栩 ; 靳辉 ; 车景辉 ; 梁淑轩 ; 李振甲 ; 林金明 ; XIN Tian-Bing ; WANG Xu ; JIN Hui ; CHE Jing-Hui ; LIANG Shu-Xuan ; LI Zhen-Jia ; LIN Jin-Ming | |
2010-05-13 ; 2010-05-13 | |
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 Chemiluminescence enzyme immunoassay,horseradish peroxidase,carcinoembryonic antigen,human serum R446.6 |
其他题名 | Microplate Chemiluminescence Enzyme Immunoassay for the Determination of Carcinoembryonic Antigen in Human Serum |
中文摘要 | 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. |
语种 | 中文 ; 中文 |
内容类型 | 期刊论文 |
源URL | [http://hdl.handle.net/123456789/31805] ![]() |
专题 | 清华大学 |
推荐引用方式 GB/T 7714 | 辛天兵,王栩,靳辉,等. 微板式化学发光酶免疫分析法测定人血清中癌胚抗原[J],2010, 2010. |
APA | 辛天兵.,王栩.,靳辉.,车景辉.,梁淑轩.,...&LIN Jin-Ming.(2010).微板式化学发光酶免疫分析法测定人血清中癌胚抗原.. |
MLA | 辛天兵,et al."微板式化学发光酶免疫分析法测定人血清中癌胚抗原".(2010). |
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