中国兽医科学2017，Vol.47Issue(7)：915-919,5.DOI:10.16656/j.issn.1673-4696.2017.07.019
鲑传染性贫血病毒TaqMan实时荧光定量RT-PCR检测方法的建立及应用
Establishment and application of a TaqMan real time RT-PCR for the detection of infectious salmon anaemia virus
摘要
Abstract
To establish a method for the rapid and quantitative detection of infectious salmon anaemia virus(ISAV),one pair of primers and TaqMan probe were designed based on NS,which is conservative and coded by segment seven ORF1.After optimizing reaction conditions,a TaqMan real time PCR for detection of ISAV was established,then the sensitivity,specificity and repeatability were estimated,and the method was used in salmon specimens which were collected between 2014 to 2016 from Chinese ports.The detection results were compared with that of the method recommended by OIE.The result showed the TaqMan method had high specificity and repeatability,and the minimum detection limit reached to 1.31× 102copies/μL.The coefficient of variation was less than l％,and 299 clinical samples detection results showed the positive detection rate of ISAV was 6.35％(19/299),which was consistent with the results by OIE,and the Ct of the method was lower than that of the method of OIE in the detection of the 19 positive specimens.This method is efficient,sensitive,specific and stable,which has broad application prospects in quarantine department,and it is meaning for the identification of ISAV.关键词
鲑传染性贫血病毒/TaqMan/荧光定量RT-PCR/口岸检疫Key words
infectious salmon anaemia virus/TaqMan/real time RT-PCR/port quarantine分类
农业科技引用本文复制引用
肖璐,王军,邬旭龙,李丹丹,杨苗,林华,陈世界,安微,王印,刘荭,严玉宝,胡娟..鲑传染性贫血病毒TaqMan实时荧光定量RT-PCR检测方法的建立及应用[J].中国兽医科学,2017,47(7):915-919,5.基金项目
四川出入境检验检疫局科技项目(SK201503) （SK201503）
