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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 a prokaryotic expression vector of toxin A and toxin B of C. difficile to obtain fusion antigen and identify its antigenic activity. Methods The genomic DNA of ATCC 43255 strain was used as a template to amplify the carboxyl-terminal (C-terminal) fragments of toxin A and B by PCR. The fragments of the carboxy-terminal toxin A and B were obtained by prokaryotic expression. The antigenicity of the toxin A and B carboxy-terminal antigenic segments was identified using a C. difficile toxin assay kit. Results The prokaryotic expression vectors of toxin A and toxin B of C. difficile were successfully constructed and the toxin A and B carboxy-terminal antigen segments recognized by the corresponding specific antibodies were obtained. Conclusion It is concluded that the toxin A and B carboxyl terminal antigens have good antigenic activity, which lays the foundation for the further preparation of the corresponding antibodies and the establishment of immunoassay for C. difficile toxin A and B.