牛传染性鼻气管炎病毒PCR检测方法的建立

Establishment and Verification of PCR for Latency of Bovine Herpesvirus 1

为建立一种牛传染性鼻气管炎病毒(IBRV)PCR检测方法,依据IBRV的t K基因序列,设计合成PCR的引物,以IBRV的标准AV20毒株为对象,确定PCR最终反应体系为25μL,并优化了PCR反应条件。结果显示,该PCR方法可以扩增出301bp的特异性片段;对牧场中常见的BVDV进行PCR特异性检测,未检测到BVDV特异性条带;10倍稀释法验证PCR优化反应后的灵敏性为可检测出0.002TCID50的病毒量。结果表明,该方法可为牧场进行IBR疫情监测提供有效技术手段。

Bovine herpesvirus 1（BHV-1）,the causative agent of infectious bovine rhinotracheitis（IBR）,is considered to be the most common viral pathogen found in bovine.PCR amplification was carried out in a final volume of 25 μL.The optimization of the PCR reaction components and cycling parameters was performed,respectively.The result of specific fragment for the standard IBRV by PCR amplified is 301 bp.The analytical specificity of the PCR was determined by testing two different types of virus were BVDV and IBRV.The sensitivity of the PCR was determined by testing sequential 10-fold dilutions of IBRV.The establishment of PCR in the assay could be an useful tool for monitoring BHV-1 in the farm.