艰难梭菌毒素A和毒素B羧基端抗原区段的表达及抗原性分析  

Expression and antigenicity analysis of C-terminal domains of Clostridium difficile toxins A and B

作　　者：黄忱[1] 杨锡琴[2] 修冰水[2] 罗芸[1] 刘志强[2] 张旭辉[2] 赵琰枫 金大智[1] 冯晓燕[2] 张贺秋[2] 

机构地区：[1]浙江省疾病预防控制中心,杭州310051 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《军事医学》2015年第12期912-914,918,共4页Military Medical Sciences

基　　金：国家自然科学基金资助项目(81471998);国家卫生计生委科学研究基金-浙江省医药卫生重大科技计划(WKJ-ZJ-1507);全军后勤科研计划资助项目(CWS13J049)

摘　　要：目的构建艰难梭菌毒素A和毒素B的原核表达载体,获得融合表达抗原,并鉴定其抗原活性。方法以ATCC43255菌株基因组DNA为模板,通过PCR扩增获得毒素A和B羧基端(C端)基因片段,并进行原核表达获得毒素A和B羧基端抗原区段。利用艰难梭菌毒素检测试剂盒对毒素A和B羧基端抗原区段的抗原性进行鉴定。结果成功构建了艰难梭菌毒素A和毒素B的原核表达载体,并获得能被相应特异性抗体所识别的毒素A和B羧基端抗原区段。结论获得毒素A和B羧基端抗原区段具有很好的抗原活性,为进一步制备相应抗体并建立艰难梭菌毒素A和B的免疫检测方法奠定了基础。Objective To construct the prokaryotic expression vector of carboxyl（ C）-terminal domains of Clostridium difficile toxins A and B,express recombinant antigens in Escherichia coli, and identify their antigenicity. Methods The genes of C-terminal domains of C. difficile toxins A and B were cloned from ATCC43255 genome DNA. The recombinant antigens were expressed in E. coli with IPTG induction and purified by Ni-NAT deads. The antigenicity was detected using C. diff Qick Chek Complete dual-antigen EIA. Results Prokaryotic expression vectors of C-terminal domains of C. difficile toxins A and B were constructed. The obtained C-terminal antigens could be identified by specific anti-toxins A and B antibodies. Conclusion The obtained C-terminal antigens of C. difficile toxins A and B can be used to prepare corresponding antibodies,which will help develop the immunoassay for C. difficile toxins A and B.

关 键 词：艰难梭菌 细菌毒素类 毒素A 毒素B 原核表达 

分 类 号：R378[医药卫生—病原生物学]