多重聚合酶链反应检测环境中产气荚膜杆菌

Multiplex PCR for Detection and Genotyping of Clostridium Perfrinens from Environmental Sample

产气荚膜杆菌根据产生 4种主要毒素 (α、β、ε、ι 毒素 )可分为 5种类型A E型。在该研究中依据产气荚膜杆菌的 5种毒素基因的序列 ,设计了 5对PCR引物 ,建立了多重PCR方法 ,该方法可以快速区分 5种类型的产气荚膜杆菌。并对 68份环境样品 (生活污水、生物肥料 )进行检测和基因分型 ,共检出 55株产气荚膜杆菌 ,其中A型产气荚膜杆菌 50株 ,占总数的 90 %以上 ,B型、C型、D型只占 5 % ,未检出E型产气荚膜杆菌 ,所有的分离株没有发现带有肠毒素。结果表明 ,目前环境中主要存在的菌型为A型菌 ,其他菌型的产气荚膜杆菌很少 。

Clostridium pertringens isolates are currenty classified into one of five biotype(A E)on the basis of the differential production of α ?β ?ε ,and ι toxin.Different biotypes associate with different diseases of man and animals.In this study a Multiplex PCR assay was developed to detect the genes encoding five major toxins(α ?β ?ε ?ι and enterotoxin). Cl.perfringens isolates taken from environmental samples,induding Sewages and bio compost were genotyped using multiplex PCR assay.Our results showed that most isolates were found to be genotype A,over 90% of total Cl.perfringens isolates,only about 5% of Cl.perfringens isolates were proved to be other types,entero toxigenic Cl.perfringens were not found in this study.