猪伪狂犬病毒野毒株套式PCR检测方法的建立和应用  

Construction and Application of a Nest PCR Assay for Identification of Field Pseudorabies Virus Strains

作　　者：张志[1] 樊雅婷[2] 吴发兴[1] 刘爽[1] 董雅琴[1] 邵卫星[1] 段纲[2] 王树双[1] 李晓成[1] 

机构地区：[1]中国动物卫生与流行病学中心,山东青岛266032 [2]云南农业大学,昆明云南650201

出　　处：《中国动物检疫》2015年第5期73-77,共5页China Animal Health Inspection

基　　金：科技基础性工作专项(2012FY111000);青岛市民生计划项目(13-1-3-91-nsh)

摘　　要：为建立能鉴别猪伪狂犬病病毒(PRV)野毒株和疫苗株的实用方法,本文以疫苗株缺失的3.5kb片段为靶基因,设计了2对PCR引物,建立了能够特异性检测PRV野毒株的套式PCR方法。与常规PCR相比,该方法检测极限可达10个病毒拷贝/μL,是常规PCR的1000倍,并与猪瘟病毒、猪蓝耳病病毒、猪细小病毒和猪圆环病毒等不存在交叉反应现象。在对临床150份样品检测时,套式PCR的阳性率为21.33%,常规PCR的阳性率为10%。这表明该方法具有特异性高、敏感性强的特点,是一种值得基层推广应用的快速诊断技术,可用于PRV的诊断和流行病学调查等。To differentiate the field and vaccine pseudorabies virus （PRV） strains, a nest PCR assay was constructed by designing two nested primers based on the 3.5 kb target gene deleted in PRV vaccine strain. The detection limit of the nest PCR assay was up to 10 virus copies/μL, without cross-action with other viruses such as classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine parvovirus and porcine circovirus, while the routine PCR detected only 104 virus copies/la L. Both the developed nest PCR assay and the routine PCR assay were used to test 150 field samples resulting in 21.33% and 10% positive rate respectively. The results indicated that this method was of high sensitivity and specificity and can be widely used for PRV diagnosis and investigation.

关 键 词：猪伪狂犬病病毒 野毒株 套式PCR 检测方法 

分 类 号：S852.65[农业科学—基础兽医学]