结核分枝杆菌三种特异性抗体ELISA检测方法的建立  

Detection of Mycobacterium Tuberculosis by ELISA Using 3 Specific Antibodies

作　　者：牛钦王[1] 李红霞[1] 曾林子[1] 徐佳楠[1] 陈建平[1] 

机构地区：[1]四川大学华西基础医学与法医学院寄生虫学教研室病原生物学研究室,成都610041

出　　处：《热带医学杂志》2010年第10期1163-1165,1186,共4页Journal of Tropical Medicine

基　　金：四川省公益性研究计划项目(No.2008SZ0106)

摘　　要：目的 建立以结核分枝杆菌特异性蛋白ESAT6、CFP10-ESAT6、ESAT6-PPE68为包被抗原检测结核病人血清中特异性抗体的间接ELISA检测方法。方法 利用本室构建并纯化的结核分枝杆菌特异性抗原蛋白ESAT6、CFP10-ESAT6、ESAT6-PPE68,建立结核分枝杆菌特异性抗体检测的间接ELISA检测方法:ESAT6-ELISA、CFP10-ESAT6-ELISA、ESAT6-PPE68-ELISA。并与PPD-ELISA同时检测220例结核病人血清、30例非结核呼吸疾病病人血清和50例健康人群血清,分别对三种方法的敏感性与特异性进行统计学分析。结果 ESAT6-ELISA的敏感性为25%,特异性为95%;CFP10-ESAT6-ELISA的敏感性为45%,特异性为93%;ESAT6-PPE68-ELISA的敏感性为48%,特异性为85%。结论 三种结核分枝杆菌特异性抗体检测方法的敏感性均低于PPD-ELISA,但三种结核分枝杆菌特异性抗体检测方法的特异性均高于PPD-ELISA,其中CFP10-ESAT6-ELISA检测的效果最佳,PPE68可作为结核特异性诊断的候选抗原。Objective Establishing indirect ELISA methods using ESAT6,CFP10-ESAT6,ESAT6-PPE68 which all are specific to Mycobacterium tuberculosis as coating antigens to detect the specific antibodies of tuberculosis patients.Methods By employing the specific antigens of Mycobacterium tuberculosis ESAT6,CFP10-ESAT6 and ESAT6-PPE68,cloned,expressed and purified by our lab,we have established three indirect ELISA methods to detect specific antibodies of Mycobacterium tuberculosis：ESAT6-ELISA,CFP10-ESAT6-ELISA and ESAT6-PPE68-ELISA.220 patients with tuberculosis,30 patients with non-tuberculosis lung disease and 50 healthy human controls were analyzed simultaneously with PPD-ELISA,and the sensitivity and the specificity of the three different methods are evaluated.Results The sensitivity of ESAT6-ELISA is 25%,with specificity of 95%;the sensitivity of CFP10-ESAT6-ELISA is 45%,with specificity of 93%;the sensitivity of ESAT6-PPE68-ELISA is 48%,with specificity of 85%.Conclusions Compared with PPD-ELISA,the sensitivity of the three ELISA methods developed by our lab lower than that of PPD-ELISA,but the specificity of the three ELISA methods developed by our lab is higher than that of PPD-ELISA,and CFP10-ESAT6-ELISA is the best.PPE68 is a specific antigen which has crucial importance for detection of tuberculosis.

关 键 词：结核分枝杆菌 ESAT6 CFP10 PPE68 ELISA 

分 类 号：R378.911[医药卫生—病原生物学]