海产品中副溶血弧菌的分离和PCR检测

Isolation and rapid detection by PCR of Vibrio parahaemolyticus from seafood

为了建立一种快速、特异的PCR方法检测副溶血弧菌(Vibrio parahaemolyticus,Vp),根据Genebank库中收录的Vp的tlh基因序列进行引物设计,应用PCR技术扩增tlh基因。利用TCBS和TSI2种选择性培养基从海产品总分离出68株疑似副溶血弧菌,经生化鉴定68株均为副溶血弧菌。用Chelex100法提取基因组DNA,进行tlh基因的PCR检测,tlh基因在不同副溶血弧菌中都广泛存在,而大肠杆菌、沙门氏菌、志贺氏菌等其他食源性致病菌均为阴性。tlh基因具有种属特异性,因此可以用来检测副溶血弧菌。

To develop a PCR assay for the rapid and specific detection of Vibrio parahaemolyticus（Vp）,primers were designed according to the gene sequences of tlh published in Genebank and PCR was applied to amplify the gene of tlh of Vibrio parahaemolyticus.Two selective media-TCBS agar and TSI were utilized for the isolation of Vibrio parahaemolyticus.According to the biochemical identication,all the 68 suspected strains which were isolated from seafood were identified to be Vibrio parahaemolyticus.The genomic DNA of Vibrio parahaemolyticus was extracted by chelex-100 and the gene of tlh of Vibrio parahaemolyticus was detected by PCR.The tlh gene is widely in Vibrio parahaemolyticus and has genus characteristics.So we can amplify the tlh gene as a rapid detection method for Vibrio parahaemolyticus.