使用二重PCR方法快速检测单核细胞增生性李斯特氏菌的研究

摘要

Two pairs of specific primers （ Hlyl -L/Hlyl-U and InlA - L/InlA - U） were designed based on conserved regions of Hly gene and InlA gene of Listeria mouocytigenes （LM） on GenBank database, and the multiplex PCR assay was established for detection of LM.The results indicated that the PCR could only amplify two specific fragments with 744bp and 520bp in length of standard strains of LM and the strains isolated from the poultry products, excluding common food-borne pathogens such as E.coli and Salmonella and the Listeria irmocua.The sensitivity of the new duplex PCR was I＆CFU/mL and it can be used to identify LM cultures isolated from the internal organs of experimentally and orally infected mice.The method showed good specificity, sensitivity and versatility for the rapid identification in detection of LM.%本研究以单核细胞增生性李斯特菌Hly基因和InlA基因作为靶序列设计2对引物，建立了检测单增李斯特菌的二重PCR方法。结果表明，该方法可以同时扩增出单增李斯特菌的Hly基因和InlA基因，而对大肠杆菌、沙门氏菌等食品中的常见致病菌和英诺克李斯特菌均不能扩增出任何条带，表现出良好的特异性。二重PCR方法的最低检测限为10^4CFU／mL，并能够用于鉴定实验小鼠攻毒后内脏器官的单增李斯特菌分离培养物。该方法表现出良好的特异性、敏感性和通用性，适用于单增李斯特菌的快速鉴别检测。