EB病毒EA优势表位区段抗原的克隆表达及其鼻咽癌诊断价值初步评价  

Cloning and Expression of Dominant Epitope Fragment Antigen of Epstein-Barr Virus Early Antigen and its Preliminary Evaluation

作　　者：陈世锋[1] 胡纪文 殷瑛 杜江东[1] 唐倩青[1] 伊茂礼[1] 王学波[1] 张玲 危利 孙成铭[1] CHEN Shi-Feng;HU Ji-Wen;YIN Ying;DU Jiang-Dong;TANG Qian-Qing;YI Mao-Li;WANG Xue-Bo;ZHANG Ling;WEI Li;SUN Cheng-Ming(Clinical Laboratory of Yantai Yuhuangding Hospital,Yantai 264000;Medical Laboratory of Shenzhen Luohu Hospital Group,Shenzhen 518001;Beijing Institute of Biotechnology,Beijing 100071;Medical Institute of Oriental Ocean(Beijing),Beijing 100071,China)

机构地区：[1]烟台毓璜顶医院检验科,山东烟台264000 [2]罗湖医院集团医学检验中心,广东深圳518001 [3]军事医学研究院生物工程研究所,北京100071 [4]东方海洋(北京)医学研究院,北京100071

出　　处：《生物技术通讯》2020年第5期505-510,共6页Letters in Biotechnology

基　　金：山东省重点研发计划(2019JZZY011018)。

摘　　要：目的:制备高活性EB病毒早期抗原(EA)的优势表位区段抗原,并初步评价该抗原在鼻咽癌诊断中的应用价值。方法:利用生物学软件分析EA抗原的B细胞表位分布,选取含有优势表位的抗原区段,然后利用分子生物学技术进行克隆表达获得纯化抗原,采用酶联免疫吸附试验(ELISA)初步评价优势表位区段抗原的检测性能。结果:选取了2个优势表位区段抗原并获得了高效表达抗原EA1(61~200 aa)和EA2(291~404 aa),经小样本验证,确定其中EA2具有更高的抗原活性和特异性。放大样本进行检测,基于EA2抗原的IgG和IgA间接ELISA方法,检测灵敏度和特异性分别为90.38%、93.68%和65.38%、95.79%。EA-IgG检测的AUC值为0.972,EA-IgA检测的AUC值为0.893。结论:优势表位区段抗原EA2可用于EA抗体检测,利用该抗原建立的EA-IgG和EA-IgA间接ELISA检测方法可以很好地区分鼻咽癌患者和健康者,EA-IgG比EA-IgA更适于鼻咽癌的鉴别诊断。Objective:The dominant epitope fragment antigen of Epstein-Barr(EB)virus early antigens(EA)were prepared and its application value in the diagnosis of nasopharyngeal carcinoma(NPC)was preliminarily evaluated.Methods:The B cell epitope distribution of EA was analyzed by BIOSUN software,and the antigen fragments containing the dominant epitope were selected.Then the dominant epitope fragment antigens were cloned and expressed using molecular biology technology,and their detection performance was preliminarily evaluated by ELISA.Results:Two antigen fragments,EA1(61~200 aa)and EA2(291~404 aa),were selected and highly expressed antigens were obtained.EA2 had higher activity and specificity than EA1.The detection sensitivities and specificities of IgG and IgA were 90.38%,93.68%,and 65.38%,95.79%,respectively,by indirect ELISA based on EA2 antigen.The AUC value of EA-IgG was 0.972 and that of EA-IgA was 0.893.Conclusion:The dominant epitope fragment antigen EA2 is the preferred antigen that can be used for EA antibody detection.The indirect ELISA detection methods of EA-IgG and EA-IgA established using EA2 can be used to distinguish NPC patients from healthy controls.EA-IgG is more suitable for differential diagnosis of nasopharyngeal carcinoma.

关 键 词：EB病毒 早期抗原 优势表位区段抗原 诊断 鼻咽癌 

分 类 号：R392.1[医药卫生—免疫学] Q78[医药卫生—基础医学]