弗尼斯弧菌PCR检测方法的建立及应用

目的:弗尼斯弧菌(Vibrio fumissii)是引起世界范围内胃肠功能紊乱的致病菌之一,一般通过摄人生的或者未煮熟的水产品进行传播。本研究旨在建立高效、可靠的PCR检测方法应用于食品中弗尼斯弧菌的快速检测。方法和结果:根据弗尼斯弧菌的toxS基因分别设计普通PCR引物及实时PCR引物、探针,建立普通PCR和实时荧光PCR方法,分别对两种方法的特异性和灵敏度进行比较分析,同时应用于384份食品样品检测。结果表明,建立的普通PCR和实时荧光PCR方法特异性良好,检测灵敏度分别为:2.4×10^3 CFU/ml和24 CFU/ml,并于5 h内完成整个检测过程,同时对384份食品进行检测,共检出6株阳性菌株。结论:本研究建立的PCR检测技术灵敏度高、特异性好,可应用于食品中弗尼斯弧菌的快速检测。

Establishment and Application of a PCR Technique Based Method for Detection of Vibrio Furnissii

Aims:Vibrio furnissii is a cause of human gastrointestinal disorders worldwide,could transmit by ingestion of raw or undercooked contaminated seafood.The aim of this work was to develop an effective and reliable method for the identification of V.furnissii.Methods and Results:Conventional PCR and real-time fluorescence PCR techniques were applied to the rapid detection of V.furnissii by designing primers and probe target for the toxS gene.The specificity and sensitivity of the method were investigated,and the practical application was analyzed using 384 commercial food products.The results showed that a high detection specificity of the developed method.The detection limit of the conventional PCR and real-time fluorescence PCR was 2.4×10^3 CFU/ml and 24 CFU/ml,respectively,and the entire identification could be completed within 5 h.In addition,the technique enabled the accurate recognition of V.furnissii from food products(6 positive products).Conclusions:A specific,sensitive and rapid PCR method target for V.furnissii toxS gene was established and could be applied to the bacterial detection in food products.