食品中产气荚膜梭菌LAMP快速检测方法的建立

摘要

LAMP (Loop-mediated isothermal amplification, LAMP) rapid detection method for Clostridium perfringens in food was established, with the specific primers designed on the CPa gene. The results of the study indicated that the method had perfect specificity, as 5 strains of Clostridium perfringens were able to amplify specific fragments, while 13 strains of non-Clostridium perfringens were not, and no false positive or false negative results occurred. The test could be done within 1 hour with the sensitivity low to 10 fg/μL. The established method provides a better choice for rapid detection of Clostridium perfringens .%应用环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP),针对产气荚膜梭菌特有的α毒素(CPa)基因序列分析设计特异性引物,建立食品中产气荚膜梭菌特异性快速检测方法.研究结果表明,所设计的引物具有良好的特异性,5株产气荚膜梭菌均能扩增出特异性片段,而13株非产气荚膜梭菌均未扩增出相应片段,无假阴性或假阳性情况出现.同时,该方法可在1 h内完成反应,且检测灵敏度达到10 fg/μL.该方法为产气荚膜梭菌的快速检测提供了一种重要的技术手段.