羊源产气荚膜梭菌不同毒素型多重PCR检测方法的建立及初步应用

发布时间：2018-04-15 23:02

本文选题：产气荚膜梭菌 + 毒素型　；参考：《中国兽医学报》2017年08期

【摘要】：根据GenBank中已发布的产气荚膜梭菌α,β,ε,ι毒素基因序列,设计并合成4对特异性引物,经过优化多重PCR反应条件,建立了检测产气荚膜梭菌不同毒素型多重PCR方法。特异性试验表明,该方法对A,B,C,D,E型产气荚膜梭菌标准菌株均扩增出了相应的目的条带,而对诺维氏梭菌和腐败梭菌扩增为阴性;灵敏性试验表明,该方法对A,B,C,D,E型标准菌株基因组DNA最低检测量分别为9.0,17.8,12.2,13.8,18.5pg;重复性试验表明,该方法有很好的重复性。应用所建立的方法从21份羊临床病料中检测出9株A型和1株C型产气荚膜梭菌。本试验建立的多重PCR方法可以进行产气荚膜梭菌的快速检测及5种毒素型的鉴别。
[Abstract]:According to the sequence of 伪, 尾, 蔚, l toxin genes of Clostridium perfringens published in GenBank, four pairs of specific primers were designed and synthesized. After optimizing the reaction conditions of multiple PCR, a method for detecting different types of multiple PCR of Clostridium perfringens was established.The specificity test showed that the target bands were amplified by this method for the standard strains of Clostridium perfringens, but negative for Clostridium norvegris and Clostridium putrefaciens, and the sensitivity test showed that,The minimum detection of genomic DNA for the standard strain of Ampelopsis Candrosemide E was 9.0 ~ 17.812.813.8 ~ 18.5pg. the reproducibility test showed that the method had good reproducibility, and the results showed that the method had good reproducibility and reproducibility, and the results showed that the method had good reproducibility and reproducibility.Nine isolates of Clostridium perfringens type A and one strain of type C were detected from 21 sheep clinical samples by using the established method.The multiplex PCR method established in this study can be used for rapid detection of Clostridium perfringens and identification of five toxin types.
【作者单位】：西北农林科技大学动物医学院;
【基金】：陕西省科技攻关资助项目(2015NY162) 西北农林科技大学试验示范站(基地)科技成果推广资助项目(TGZX2015-32)
【分类号】：S852.61

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