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癌抗原125化学发光免疫定量分析方法的建立 预览

Establishment of a chemiluminescent immunoassay for cancer antigen 125
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摘要 目的建立癌抗原125(cancer antigen125,CA125)的化学发光免疫定量分析方法。方法采用双抗体异位点一步夹心法,以CA125单克隆抗体作为固相包被,采用改良的过碘酸钠法进行CA125多克隆抗体与碱性磷酸酶偶联制备酶标抗体,并优化标记条件;以金刚烷胺衍生物(CSPD)及增强剂(SapphireⅡ)作为底物;用国家标准品校定定量标准品,建立了人血清CA125的化学发光免疫定量分析法。用该法检测500份正常人血清,确定正常值参考范围,将临床血清标本126份与BayerACS:180全自动发光体系统进行比较。结果本方法的最低检出量为0.1U/ml,特异性100%,线性范围0.1—960.0U/ml,检测内和检测间的变异系数分别为3.6%和5.3%,回收率在86.5%-96.2%之间。与CA50的交叉≤0.1%,与CA153≤0.1%,与CA199≤0.5%,与CEA≤0.1%,与BayerACS:180相关系数r=0.993(P〈0.001)。结论建立的CA125化学发光免疫定量分析法结果可靠,可用于临床检测。 Objective To establish a chemiluminescent immunoassay for determining cancer antigen 125 (CA125)in human serum. Methods Using two-antibodies sandwich enzyme immunoassay, one monoclonal antibody was coated to a microplate, and a polyclonal antibody was conjugated with alkaline phosphatase. In the same time, the conjugation was performed by optimized NalQ method. Chemi- luminescent immunoassay was established using CSPD and Sapphire 11 luminescent system. Sensitivity, specificity,linear scope, precision, interference and recover), for the method were evaluated. The reference range was defined by testing 500 serum samples of healthy subjects. The data of 126 serum samples of patients were assayed by chemiluminescent immunoassay, and the relation with the results by the Bayer ACS:180 ( automated immunoassay system) was analyzed. Results The sensitivity was 0. 1 U/ml. The specificity was 100%. The linear scope was 0.1 - 960.0 U/ml. Average inter and intra precisions were 3.6% and 5.3%, respectively. The recoveries ranged from 86.5% to 96.2%. The cross-reacting rates for CAS0,CA153 ,CA199,CEA were≤0.1% ,0.1% ,0.5% and 0.1% ,respectively. The results assayed by chemiluminescent immunoassay was positively related to the results by ACS: 180 kit of Bayer Co. (r = 0. 993 ,P 〈0.001 ). Conclusion A successfully established chemiluminescent immunoassay is a feasible way for clinically determining CA125 with dependable results.
作者 王克珍 买制刚 蒋宏伟 丁新荣 WANG Ke-zhen , MAI Zhi-gang , JIANG Hong-wei , DING Xin-rong ( R1A Center, Shaanxi Provincial People' s Hospital, Xi' an 710068, China ; 2 Biotechnology Center of Shenzhen University)
出处 《山西医科大学学报》 CAS 2009年第12期 1077-1079,共3页 Journal of Shanxi Medical University
基金 陕西省科技社发攻关计划基金资助项目(2007K09-09)
关键词 癌抗原125 化学发光免疫法 碱性磷酸酶 cancer antigen 125 chemilumineseent immunoassay alkaline phosphatase
作者简介 王克珍,女,1963—01生,硕士,副主任技师.
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