牛病毒性腹泻病毒实时荧光定量RT-PCR方法的建立及应用  

Establishment and Application of Real-time Fluorescent Quantitative RT-PCR Method for Detection of Bovine Viral Diarrhea Virus

作　　者：王荣[1] 李文文[1] 王研[1] 刘亚刚[1] 余琼[2] 任永刚[2] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]西昌市畜牧局,四川西昌615000

出　　处：《中国畜牧兽医》2014年第2期35-39,共5页China Animal Husbandry & Veterinary Medicine

基　　金：西南民族大学硕士研究生创新型科研项目(CX2013SZ71)

摘　　要：持续性感染和免疫耐受是牛病毒性腹泻病的重要特征,这一特征给该病的诊断、检疫及防控造成很大困难,为此,建立快速实用、高效敏感的牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)检测方法具有重要意义。据GenBank中登录的BVDV 5′UTR保守区域设计并合成1对特异性引物,以SYBR GreenⅠ染料为扩增指示剂,建立了BVDV实时荧光定量RT-PCR检测方法。结果表明,该方法具有快速、敏感、特异、重复性好等优点。该方法的建立为牛病毒性腹泻病的早期快速诊断和有效检出持续性感染动物提供了手段,是该病检疫和诊断方法的补充和完善。Persistent infection and immunologic tolerance are important characteristics of bovine viral diarrhea disease, which have caused great difficulties on the diagnosis, quarantine and prevention of the bovine viral diarrhea virus （BVDV）, so a fast and efficient detection method of BVDV is quite necessary. A pair of primers were designed based on the gene sequences of BVDV 5rUTR published in GenBank, then established the Real-time fluorescent quantitative RT-PCR detection method with SYBR Green I to detect BVDV. The results showed that the detection method had high specificity, strong sensitivity and good repeatability. Thus, the detection method of the Real-time fluorescent quantitative RT-PCR would provide an effective means for the rapid detection of BVDV and persistent infected animals. It could supplement and perfect the quarantine and diag- nostic methods of BVDV.

关 键 词：牛病毒性腹泻病毒 实时荧光定量RT—PCR 5′UTR 检测方法 

分 类 号：S852.659[农业科学—基础兽医学]