中国兽医科学2017，Vol.47Issue(9)：1124-1128,5.DOI:10.16656/j.issn.1673-4696.2017.09.009
蓝舌病病毒和鹿流行性出血热病毒双重荧光定量RT-PCR检测方法的建立
Establishment of a duplex real-time RT-PCR for simultaneous detection of BTV and EHDV
摘要
Abstract
To develop a method for simultaneous detection of Bluetongue disease virus(BTV) and Epizootic hemorrhagic disease virus(EHDV),a duplex real-time RT-PCR was established with the specific primers and TaqMan fluorescent probes designed according to the conserved gene sequences of BTV and EHDV.The reaction conditions,specificity and sensitivity of this method were optimized.Clinical samples were collected for verification of this method.The results showed that the assay was highly specific and sensitive with the detection limit of 1×102 copies/μL for BTV and EHDV,but no amplification for FMDV,VSV,SBV,PPRV,GTPV and SPPV.In addition,the coefficient of variation was less than 2.55％ for both intra-assay and inter-assay.Moreover,clinical test showed that the method could accurately detect the nucleic acids of BTV and EHDV in clinical tissues,It is indicated that the duplex real-time RT-PCR can be used as an effective tool for rapid detection and epidemic surveillance of BTV and EHDV.关键词
蓝舌病病毒/鹿流行性出血热病毒/双重荧光定量RT-PCRKey words
bluetongue disease virus/epizootic hemorrhagic disease virus/duplex real-time RT-PCR分类
农业科技引用本文复制引用
刘佳佳,王慧煜,艾军,韩雪清..蓝舌病病毒和鹿流行性出血热病毒双重荧光定量RT-PCR检测方法的建立[J].中国兽医科学,2017,47(9):1124-1128,5.基金项目
“十二五”国家科技支撑计划项目(2013BAD12B03) （2013BAD12B03）
