Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood

查看全文 作　　者：Bin [1]Liu;Xiaohua [1]He;Wanyi [1]Chen;Shuijing [1]Yu;Chunlei [1]Shi;Xiujuan [1]Zhou;Jing [1]Chen;Dapeng [1]Wang;Xianming [1]Shi

机构地区：[1]MOST-USDA Joint Research Center for Food Safety and Bor Luh Food Safety Center,School of Agriculture and Biology,Shanghai Jiao Tong University,Shanghai 200240,China

出　　处：《Protein & Cell》索引2012年第3卷第3期,共9页

基　　金：supported by grants from the Ministry of Sci-ence and Technology of China(Nos.2012AA101601 and 2011DFA31220);the National Natural Science Foundation of China(Grant Nos.31171690,30972485,31000779 and U1031003);the Science and Technology Commission of Shanghai Municipality(Nos.10DZ0503500 and 10142201300). 摘　　要：A real time PCR assay for the detection of Vibrio para-haemolyticus in seafood samples was developed using a novel specific target and a competitive internal ampli-fication control(IAC).The specificity of this assay was evaluated using 390 bacterial strains including V.parahaemolyticus,and other strains belonging to Vibrio and non-Vibrio species.The real time PCR assay un-ambiguously distinguished V.parahaemolyticus with a detection sensitivity of 4.8 fg per PCR with purified genomic DNA or 1 CFU per reaction by counting V.parahaemolyticus colonies.The assays of avoiding interference demonstrated that,even in the presence of 2.1μg genomic DNA or 107 CFU background bacteria,V.parahaemolyticus could still be accurately detected.In addition,the IAC was used to indicate false-negative results,and lower than 94 copies of IAC per reaction had no influence on the detection limit.Ninety-six sea-food samples were tested,of which 58(60.4%)were positive,including 3 false negative results.Conse-quently,the real time PCR assay is effective for the rapid detection of V.parahaemotyticus contaminants in seafood. 关键词：Vibrio parahaemolyticus real time PCR internal amplification control SEAFOOD

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Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood

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