聚合酶链反应鉴定艰难梭菌产毒株

Identiflcation of toxigenic clostridium difrlcille by polymerasc chain reaction

以艰难梭菌Ａ毒素基因非重复片段中的２个寡核苷酸链作引物，扩增３０６ｂｐ，用多聚酶链反应技术扩增３１株细菌，结果１９株艰难梭菌产毒株均扩增出单一特异的电泳带，而８株艰难梭菌无毒株、２株索氏梭菌和２株大肠杆菌均无特异带出现。将艰难梭菌产毒株的模板ＤＮＡ从５０ｎｇ稀释至０．５ｎｇ后再进行聚合酶链反应，结果仍可见特异扩增带。说明聚合酶链反应鉴定艰难梭菌产毒株较之细菌分离培养、细胞毒素测定方法具有快速、简便、特异、敏感的优点，可望用于临床标本的直接检测。

olymerase chain reaction was used for the identification of toxigenic Clostridium difficille A primer pair derived froin non一repeating sequences of the toxin A gene were used to amplify 306bp DNA fragments.Amplified products were visualized by polyacrylmide gel electrophoresis followed by ethidium bromide staining All 19 strains of toxigenic Clostridium difficile generated single specific amplified DNA In contrast,none of the 8 strains of non一toxigenic Clostridium difficille,2 strains of Clostridium sordelli and 2 strains of E.coli gave positive results.After the detected DNA of toxigenic Clostridium difficille was diluted to 0.5ng,the polymerase chain reaction assays were still positive. The results demonstrated that polymerase chain reaction is a simple,rapid,specific and sensitive method for the identification of toxigenic Clostridium difficille which would be used for the direct detection of Clostridium difficille in feces sample.