首页|期刊导航|中国海洋湖沼学报（英文版）|A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda
中国海洋湖沼学报（英文版）2012，Vol.30Issue(5)：731-737,7.
A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda
A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda
摘要
Abstract
Edwardsiella tarda has become one of the most important emerging pathogens in aquaculture industry.Therefore,a rapid,reproducible,and sensitive method for detection and quantification of this pathogen is needed urgently.To achieve this purpose,we developed a TaqMan-based real-time PCR assay for detection and quantification of E.larda.The assay targets the hemolysin activator HlyB domain protein of E.tarda.Our optimized TaqMan assay is capable of detecting as little as 40 fg of genomic DNA per reaction.A standard curve was generated from the threshold cycle values(y)against log10(E.tarda genomic DNA concentration)as x.The intra-and inter-assay coefficient of variation(CV)values were less than 2.06％ and 1.05％ respectively,indicating that the assay had good reproducibility.This method is highly specific to E.tarda strains,as it shows no cross-reactivity to Edwardsiella ictaluri,a member of the same genus,or to nine other fish-pathogenic bacteria species belonging to three other genera.This sensitive and specific real-time PCR assay provides a valuable tool for diagnostic quantitation of E.tarda in clinical samples.关键词
Edwardsiella tarda/TaqMan/real-time PCR/detectionKey words
Edwardsiella tarda/TaqMan/real-time PCR/detection引用本文复制引用
XIE Guosi,HUANG Jie,ZHANG Qingli,HAN Nana,SHI Chengyin,WANG Xiuhua,LIU Qinghui..A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda[J].中国海洋湖沼学报（英文版）,2012,30(5):731-737,7.基金项目
Supported by the Special Fund for Agro-scientific Research in the Public Interest(No.201103034)and the Construction Special Fund of Modern Agriculture and Industrial Technology Research System(No.CARS-47) （No.201103034）
