中国兽医科学2017，Vol.47Issue(6)：750-754,5.DOI:10.16656/j.issn.1673-4696.2017.06.013
牛轮状病毒实时荧光定量PCR检测方法的建立
Development of real-time PCR for detection of bovine rotavirus
摘要
Abstract
To establish a rapid and specific real-time fluorescent quantitative PCR assay to detect bovine rotavirus,the VP6 gene fragment of bovine rotavirus NCDV strain(211 bp) was cloned into the pMD19-T vector as the standard of recombinant plasmid and a SYBR Green Ⅰ fluorescent quantitative PCR detection method was established.The results showed that the recombinant plasmid containing 211 bp length VP6 gene was successfully constructed and the minimum detectable amount of this method was 15.39 copies/μ L.The results for detection of TGEV,BPV,PEDV and PRV by this method were negative,which showed that this method had good specificity.The results of repetition experiment showed the coefficient of variation of intra-assay and inter-assay were less than 3％,which showed that this method had good repeatability.In the present study,the qRT-PCR detection method basing on VP6 gene of bovine rotavirus was successfully established.This method is specific,sensitive and rapid,and can be used for the early diagnosis of bovine rotavirus.关键词
牛轮状病毒/VP6基因/实时荧光定量PCRKey words
bovine rotavirus/VP6 gene/real-time fluorescence quantitative PCR分类
农业科技引用本文复制引用
王梓,乔薪瑗,刘佳丽,赵莉,王瑞翀,李一经,王丽,唐丽杰,徐义刚,刘敏..牛轮状病毒实时荧光定量PCR检测方法的建立[J].中国兽医科学,2017,47(6):750-754,5.基金项目
“十三五”国家重点研发计划项目(2016YFD0500904) （2016YFD0500904）
