中国兽医科学2017，Vol.47Issue(12)：1481-1485,5.DOI:10.16656/j.issn.1673-4696.2017.12.002
猪环曲病毒TaqMan荧光定量RT-PCR检测方法的建立和应用
Development and application of the TaqMan real-time reverse transcription polymerase chain reaction for detection of porcine torovirus
摘要
Abstract
The real-time RT-PCR assay was developed using the specific primers and a TaqMan probe according to the N gene conserved sequence of porcine torovirus.Specificity,sensitivity and repeatability of the method were evaluated.The result showed this method had no cross reactions with other diarrhea virus(porcine epidemic diarrhea virus,transmissible gastroenteritis virus,porcine astrovirus,group A porcine rotavirus,porcine bocavirus,porcine kobuvirus,porcine pseudorabies virus).The Ct value of standard curve had a linear relationship to the log of the final concentration of template in the range of 109～102 copies/μL.The detection limit was 100 copies/μL.The coefficient of variation was less than 2.5％ both for intra and inter assay.Among the 88 fecal samples from porcine,30 and 28 were positive based on the TaqMan real-time PCR and conventional RT-PCR methods,respectively,and the total concordance of the two methods was approximately 93.34％.In conclusion,the real-time RT-PCR could be used for the detection and the epidemiological investigation of porcine torovirus.关键词
猪环曲病毒/荧光定量RT-PCR/TaqManKey words
porcine torovirus/TaqMan real-time RT-PCR/TaqMan分类
农业科技引用本文复制引用
谢荣辉,刘霞,赵灵燕,黄靖,柴娟,吴赟竑,徐辉..猪环曲病毒TaqMan荧光定量RT-PCR检测方法的建立和应用[J].中国兽医科学,2017,47(12):1481-1485,5.基金项目
浙江省公益技术研究项目(2016C32053) （2016C32053）
