CFP21-ELISPOT方法的建立及其在结核分枝杆菌感染诊断中的应用  

The CFP21 protein-ELISPOT method of diagnosing Mycobacterium tuberculosis infection and its diagnostic value

作　　者：谢富佳[1,2] 谭继英[1,3] 乔梅[1,2] 梁正羽[1,2] 王世婷[4] 唐克峰[1,3] 祝秉东[1] 杨燕[1] 章国平[1] 

机构地区：[1]兰州大学结核病研究中心,甘肃兰州730000 [2]兰州大学第一临床医学院,甘肃兰州730000 [3]兰州大学基础医学院免疫学研究所,甘肃兰州730000 [4]兰州市第二人民医院检验科

出　　处：《中国病原生物学杂志》2011年第9期647-649,683,共4页Journal of Pathogen Biology

基　　金：卫生部艾滋病和病毒性肝炎等重大传染病防治科技重大专项(No.2008zx100030135);教育部"国家大学生创新性实验计划"项目(No.081073017);中央高校基本科研业务费专项资金"重点项目(No.lzujbky-2010-k07)

摘　　要：目的用酶联免疫斑点试验(enzyme-linked immunospot assay,ELISPOT)评价检测CFP21蛋白在结核分枝杆菌感染中的诊断效果,同时,比较CFP21、CFP10和ESAT6的应用价值。方法取28例临床确诊结核患者和26例健康志愿者外周血,分离单个核细胞(peripheral blood mononuclear cells,PBMCs),用CFP21、CFP10和ESAT6蛋白刺激,定量检测PBMC分泌的特异性γ干扰素(IFN-γ)水平,判断结核分枝杆菌感染情况。用ELISA检测血清抗结核抗体。结果 ELISA检测结核病患者血清结核抗体阳性率为57.1%;以CFP21、CFP10和ESAT6为抗原,用ELISPOT方法检测结核病患者IFN-γ阳性率分别为25.0%、39.3%和32.1%。结论应用CFP21蛋白作为抗原建立的ELISPOT方法有潜在的临床应用价值,但CFP21作为抗原用于结核分枝杆菌感染诊断的敏感性低于CFP10和ESAT6。Objective To detect tuberculosis infection by enzyme-linked immunospot（ELISPOT） assay and evaluate the value of using the CFP21 protein to diagnose tuberculosis（TB） in comparison to CFP10 and ESAT6.Methods Peripheral blood mononuclear cells（PBMCs） isolated from patients with tuberculosis（n=28） and healthy volunteers serving as controls（n=26） were co-cultured with CFP21,CFP10,and ESAT6 antigens.The levels of IFN-γ were evaluated.Anti-tuberculosis antibodies were detected using ELISA.Results The 28 patients with TB tested positive for antibodies at a rate of 57.1% according to ELISA.Patients tested positive for CFP21,CFP10,and ESAT6 antigens at rates of 25.0%,39.3%,and 32.1%,respectively.Conclusion CFP21 antigen has potential for clinical use but it was less sensitive at diagnosing Mycobacterium tuberculosis infection than CFP10 and ESAT6 antigens.

关 键 词：酶联免疫斑点试验(ELISPOT) CFP21蛋白 CFP10蛋白 ESAT6蛋白 结核病 诊断 

分 类 号：R378.911[医药卫生—病原生物学]