中国动物传染病学报2016，Vol.24Issue(1)：44-51,8.
水产品中霍乱弧菌、副溶血弧菌和创伤弧菌多重PCR检测方法的建立
DEVELOPMENT OF A MULTIPLEX POLYMERASE CHAIN REACTION ASSAY FOR SIMULTANEOUS DETECTION OF VIBRIOPARAHAEMOLYTICUS, VIBRIOCHOLERAE AND VIBRIOVULNIFICUS IN AQUATIC FOODS
摘要
Abstract
Vibriocholerae (Vc), Vibrioparahaemolyticus (Vp) and Vibriovulnificus (Vv) are of major concerns in aquatic food industry as they are important pathogens causing food-borne diseases throughout the world. Therefore, it is very necessary to develop an effective multiplex polymerase chain reaction (PCR) for the simultaneous detection of these important Vibrio species. The groEL gene is a potential marker for simultaneous detection of these species in seafood samples. Three sets of species-specific primers were designed to develop a multiplex PCR. The reaction conditions of the multiplex PCR were optimized to establish a rapid, sensitive and convenient PCR method. The results showed that the multiplex PCR produced amplicons with expected sizes of 418 bp, 644 bp and 192 bp specific for Vc, Vp and Vv, respectively. This PCR method showed good efficiency as it detected three species in the same samples. The detection limits were demonstrated to be 102CFU/mL for Vp and Vc and 103CFU/mL for Vv in the mixed DNA samples. In addition, the multiplex PCR specifically amplified target species but did not cross-react with other Vibrio and non-Vibrio species. The results indicated that the multiplex PCR was a quick and cost-efficient method for detection of Vibrio species in seafoods and for epidemiological investigation.关键词
霍乱弧菌/副溶血弧菌/创伤弧菌/groEL基因/多重PCRKey words
Vibriocholerae/Vibrioparahaemolyticus/Vibriovulnificus/groELgene/Multiplex PCR分类
农业科技引用本文复制引用
蒋蔚,易力,陈永军,何再平,白雪瑞,赵梦苏,刘景云,王权..水产品中霍乱弧菌、副溶血弧菌和创伤弧菌多重PCR检测方法的建立[J].中国动物传染病学报,2016,24(1):44-51,8.基金项目
上海市科学技术委员会科研计划项目（13DZ0502702）；公益性行业(农业)科研专项 （农业）
