实时荧光定量RT-PCR检测猪水疱病病毒的研究  

Detection of Swine vesicular disease virus by Real-time,Fluorogenic Taq Man Reverse Transcription Polymerase Chain Reaction Assay

作　　者：钟金栋[1] 花群义[2] 肖荣海[2] 夏雪山[1] 杨云庆[2] 周晓黎[2] 董俊[2] 邓祖洪 

机构地区：[1]昆明理工大学生化学院,云南昆明650224 [2]云南出入境检验检疫局技术中心,云南昆明650228 [3]西双版纳兽医站,云南景洪666100

出　　处：《动物医学进展》2006年第8期61-66,共6页Progress In Veterinary Medicine

基　　金：国家"十五"重大科技攻关项目(2001BA804A48)和(2004BA519A40)

摘　　要：按照猪水疱病病毒结构蛋白VP1基因序列,设计合成引物和探针,经各反应条件的优化,建立了实时荧光定量RT-PCR技术,对猪水疱病病毒进行了特异性、敏感性和重复性试验。结果表明,实时荧光TaqManRT-PCR可检测到每个反应相当于1×102拷贝病毒RNA;与FMDV、VSV等其他水疱性病毒不发生交叉反应;制作的标准曲线中各浓度范围内有极好的线性关系且线性范围宽,相关系数为0.993;与常规的RT-PCR比较,该方法具有快速、特异、敏感、重复性好、可同时检测大量样品等优点。A fluorogenic TaqMan RT-PCR assay using a primer/probe set designed from the VP1 region of the virus genome was developed for the specificity and sensitivity detection of swine vesicular disease virus. The number of genome equivalents which were detected 1 × 10^2 copies per reactions. No cross-reactivity was demonstrated when this primer/probe sets were tested with RNA prepared from FMDV and VSV. the relationship between the values of threshold cycle（Ct）and the copy number was linear（r=0. 993） . The fluorogenic RT-PCR provides relatively fast results, greater sensitivity, enables quantitative assessment to be made of virus amounts and can handle more samples and/or replicates of samples in a single assay than the conventional RT-PCR procedure.

关 键 词：猪水疱病病毒 荧光TaqMan逆转录 聚合酶链反应 检测 

分 类 号：S852.659.1[农业科学—基础兽医学] Q503[农业科学—兽医学]