中国兽医科学2017，Vol.47Issue(4)：442-447,6.DOI:10.16656/j.issn.1673-4696.2017.04.006
水泡性口炎病毒双重荧光RT-PCR鉴别检测方法的建立
Development of a duplex real-time RT-PCR assay for the detection of vesicular stomatitis virus
摘要
Abstract
To establish a rapid,sensitive and specific assay for simultaneous detection and differentiation of vesicular stomatitis virus(VSV),a duplex real-time fluorescent RT-PCR assay was developed with two pairs and two probes of primers designed according to the L gene of vesicular stomatitis Indiana virus(VSV-IND) and vesicular stomatitis New Jersey virus(VSV-NJ).Under the optimized reaction conditions,VSV-IND and VSV-NJ were specifically detected simultaneously by the assay without cross-reaction with other inactivated virus antigens,including FMDV,BTV,BVDV,SVDV,CSFV and PRRSV RNA.Compared with the conventional RT-PCR,the assay was fast and sensitive.Moreover this assay had good reproducibility,with the coefficient of variation of Ct values less than 3％.A total of 126 clinical samples were tested by the established assay and conventional RT-PCR,and the test results were consistent.In conclusion,the established assay is rapid,specific and sensitive,and is useful for detection of VSV-IND and VSV-NJ simultaneously.关键词
水泡性口炎病毒/双重荧光RT-PCR/鉴别检测Key words
vesicular stomatitis virus/duplex real-time fluorescent RT-PCR/detection分类
农业科技引用本文复制引用
林彦星,陶虹,廖立珊,花群义,曹琛福,刘建利,张彩虹,吕建强,杨俊兴,孙洁,林庆燕,阮周曦..水泡性口炎病毒双重荧光RT-PCR鉴别检测方法的建立[J].中国兽医科学,2017,47(4):442-447,6.基金项目
国家科技支撑计划项目(2013BAD12B03) （2013BAD12B03）
“十三五”国家重点研发计划课题项目(2016YFC1200904) （2016YFC1200904）
