C型产气荚膜梭菌PCR快速检测方法的建立

Establishing rapid detection of type C Clostridium perfringens by PCR

为了对产气荚膜梭菌进行快速检测,本研究以GenBank发表的产气荚膜梭菌α毒素、β毒素作为参考,应用生物学软件设计扩增C型产气荚膜梭菌α毒素和β毒素的特异性引物,预期基因片段大小分别为1 110 bp与927 bp;建立PCR反应体系并设置反应条件;反应结束后对扩增产物进行琼脂糖凝胶电泳进行检测。结果表明,设计的引物可以良好的扩增出C型产气荚膜梭菌的α毒素和β毒素基因,扩增的基因片段大小与预期的一致,阴性水对照与ε毒素对照扩增电泳泳道均为阴性。此方法用于判定C型产气荚膜梭菌的毒素型是可行的。

According to reference sequence of α toxin and β toxin in GenBank,α toxin and β toxin primers of Clostridium perfringens type C were designed by biology software and expected gene fragment size of α toxin and β toxin were 1 110 bp and 927 bp.PCR reaction system and the reaction conditions were established,and amplified products were detected by agarose gel electrophoresis when the PCR reaction end.The results showed that the primers could amplify α toxin and β toxin genes of Clostridium perfringens type C and genes fragments were consistent with the expected size,and the negative water control and ε toxin control were negative.Therefore,the rapid PCR detection method for Clostridium perfringens type C can provide detection basis for toxin type of Clostridium perfringens.