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

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.