多重PCR鉴定不同毒素型的产气荚膜梭菌菌落.PDF

微生物学通报 JUN 20, 2008, 35(6): 989~993 Microbiology © 2008 by Institute of Microbiology, CAS tongbao@ 生物实验室 多重PCR 鉴定不同毒素型的产气荚膜梭菌菌落 * 赵 耘 杜昕波 李伟杰 康 凯 陈 敏 ( 100081) 摘 要: 参照文献报道的产气荚膜梭菌α, β, ε, τ 毒素基因cpa 、cpb 、etx 及iA 序列合成了针对4 种毒素基因的4 对特异引物, 建立了一种简单的产气荚膜梭菌定型的菌落多重PCR 方法。结果本 所保存的A, B, C, D, E 各型产气荚膜梭菌参考菌株均扩增出了相应的预期条带, 而诺维氏梭菌、 腐败梭菌和破伤风梭菌的扩增均为阴性; 将单个菌落稀释 100 倍利用此菌落多重PCR 仍能扩增到 相应的目的片段。并利用此多重 PCR 对 13 株不同动物来源的产气荚膜梭菌进行了定型鉴定, 并 与毒素中和试验鉴定结果进行了比较, 结果表明两种方法具有较高的符合率。本方法的建立对于 产气荚膜梭菌的快速检测、定型具有十分重要的意义。 关键词: 产气荚膜梭菌, 菌落多重PCR, 定型, 毒素 Toxintyping of Clostridium perfringens Strains by Colony Multiplex PCR * ZHAO Yun DU Xin-Bo LI Wei-Jie KANG Kai CHEN Min (China Institute of Veterinary Drug Control, Beijing 100081) Abstract: Four primers against the genes encoding (cpa, cpb, etx, and iA) four major toxins(α, β, ε, ι) of Cl. perfringens were designed and the colony multiplex PCR of identification and genotyping of Cl. perfringens strains were developed. Cl. perfringens reference strains stored in china institute of veterinary drug control including A, B, C, D and E genotyping were genotyped using the colony muitiplex PCR assay. The expected sequences were obtained successfully by the colony multiplex PCR assay. But the sequences were not ob- tained from Cl. novyi, Cl.