基于lolB和toxR基因的水产品中霍乱弧菌双重PCR检测方法  

Detection of Vibrio cholerae from Aquatic Animals by Dulplex PCR Based on lolB and toxR Genes

作　　者：秦国民[1] 张晓君[1] 毕可然[1] 阎斌伦[1] 秦蕾[1] 

机构地区：[1]淮海工学院海洋学院,江苏省海洋生物技术重点建设实验室,江苏连云港222005

出　　处：《食品科学》2011年第22期273-275,共3页Food Science

基　　金：江苏省水产三项工程项目(PJ2010-58);江苏省六大人才高峰项目(2009);江苏省自然科学基金项目(BK2009163)

摘　　要：选择lolB和toxR两种基因序列设计2对特异性引物,建立一种针对霍乱弧菌所有生物型菌株的双重PCR检测方法,扩增目的片段大小分别为519bp和779bp。结果表明:在同步扩增中,仅霍乱弧菌模板可同时扩增出2种基因片段,4株对照菌模板无任何扩增条带;敏感性检测结果显示,该双重PCR最低能检测3.42×103CFU/mL菌落数的霍乱弧菌。所建立的基于lolB和toxR两种基因的双重PCR检测方法特异性强、敏感性高、方法简单、用时短,可用于霍乱弧菌检测。V.cholerae is well recognized as pathogenic bacteria that cause diseases in both human and aquatic animals.In this study,two pairs of specific primers were designed based on the lolB and toxR genes,and a dulplex polymerase chain reaction（PCR） was established to detect all V.cholerae serogroup and biotypes.The results indicated that both PCR primer pairs could simultaneously amplify 519 bp and 779 bp gene fragments from the chromosomal DNA of V.cholerae.No positive reaction was detected in 4 control strains of pathogenic Vibrio.The sensitivity of the dulplex PCR assay also showed that the designed primer pairs could detect V.cholerae at a level of 3.42 × 103 CFU/mL.The assay was simple,rapid,specific,sensitive and applicable for the detection of V.cholerae for different purposes.

关 键 词：霍乱弧菌 lolB基因 toxR基因 双重PCR 

分 类 号：S941[农业科学—水产养殖]