摘要
为建立检测口蹄疫病毒(FMDV)的方法,本研究根据Gen Bank中FMDV的2B基因序列,设计合成一对引物和一条Taq Man探针,将2B基因克隆到p Blue Script SK(-)载体中,利用T7体外转录试剂盒制备标准品,通过优化反应条件,建立了Taq Man荧光定量PCR检测方法。结果表明,该检测方法的敏感性达到102拷贝/μL;与其它主要相关病毒均不发生交叉反应,批内和批间试验重复性的变异系数(CV)均小于3%。本研究建立的FMDV Taq Man荧光定量PCR方法对FMDV的快速检测具有重要意义。
In order to establish a sensitive method for foot and mouth disease virus （FMDV） detection,a TaqMan real-time PCR assay was developed using a pair of primers and specific probe designed according to the conserved region of FMDV 2B gene,which was cloned into pBlueScriptSK （-） vector to prepare 2B RNA transcribed by T7 in vitro transcription kit for setting up the standard curve.Under the optimized reaction conditions,the test results showed that this method was specific for detecting FMDV with a detection limit of 102 copies/μL,and no cross-reactions to other swine viruses.The repeatability tests indicated that the inter-and intra-variation were less than 3％.The development of this rapid and sensitive detection method could be applied in clinical diagnosis of FMDV infections and quantitative analysis of FMDV.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2014年第12期948-951,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家国际科技合作项目(2010DFB33620)
