【作者】 周姣； 刘宝生； 李昆； 陈瑞光； 张锦华；

【Author】 Jiao zhou;Kun Li;Baosheng Liu;Ruiguang Chen;Jinhua Zhang;College of Animal Science and Technology Jiangxi Agricultural University;

【机构】 江西农业大学动科院；

【摘要】 本试验根据产气荚膜梭菌α、β、β2、ε、τ和e毒素基因分别设计合成6对引物,建立PCR检测方法,对江西部分地区56份仔猪腹泻样品进行肠毒素基因检测,结果检测出2份e毒素基因阳性的样品。采用改良D-环丝氨酸培养基对54份e毒素基因阴性的样品进行产气荚膜梭菌分离,得到33株产气荚膜梭菌分离株。然后建立多重PCR(α、β、β2、ε)和τ毒素PCR方法对33株分离进行检测,结果有93.9%(31/33)菌株属于A型且携带beta2毒素。所扩增的beta2毒素与e毒素片段大小分别为566bp、516bp,与预期相一致;目的片段经克隆测序后,与产气荚膜梭菌参考株beta2毒素与e毒素基因序列的核苷酸同源性分别为99%~100%、98%~100%,所分离菌株τ毒素基因PCR检测均为阴性。更多还原

【Abstract】 In this study,Six primers are synthetized respectively according to the alpha,beta,beta 2,epsilon,t and enterotoxin gene of Clostridium perfringens in order to set up a detection method of PCR.Using this method,we got two enterotoxin gene positive samples in 56 samples of diarrhea piglets from parts of Jiangxi province.Then 33 C.perfringens are isolated from the rest of 54 enterotoxin gene negative samples using the modified medium containing D-ring serine.A multiplex polymerase chain reaction assay(alpha,beta,beta 2,epsilon) and a simple PCR about t toxin are established to test the toxin genotyping of 33 strains.the results showed that 93.9%(31/33) strains belonged to type A carrying beta2 toxin gene.The size of beta2 toxin and enterotoxin fragment is 566 bp and 516 bp respectively.They both are consistent with the expected.Sequence analyses indicated that the beta 2 toxin and enterotoxin gene sequences of 31 Jiangxi strains showed homologies similarity 99%~ 100%and 100%~ 98%respectively to standard strains of Clostridium perfringens.All 31 isolates are negative in t toxin genes after detecting by the method of PCR.更多还原