猪博卡病毒实时荧光定量PCR检测方法的建立  

Development of a Real-time PCR for Detection of Porcine Bocavirus

作　　者：邓波[1] 刘佩红[1] 周锦萍[1] 王建[1] 刘健[1] 鞠厚斌[1] 葛菲菲[1] 崔立[2] 华修国[2] 

机构地区：[1]上海市动物疫病预防控制中心,上海201103 [2]上海交通大学上海市兽医生物技术重点实验室,上海200240

出　　处：《动物医学进展》2012年第9期70-74,共5页Progress In Veterinary Medicine

摘　　要：根据GenBank公布的猪博卡病毒(PBoV)序列,通过VP1/2基因设计引物和Taq Man探针建立实时荧光定量PCR检测方法。建立的方法与PPV、PRRSV及PCV均无交叉反应,具有较高特异性,在107 copies/mL～101 copies/mL模板范围内具有良好的线性关系,所制作的标准曲线相关系数为0.997,最低可检测到101copies/mL的阳性质粒。说明所建立的PBoV实时荧光定量PCR检测方法具有灵敏度高、特异性好和精确性高等优点。A TaqMan based real-time PCR was developed for the specific detection of porcine bocavirus virus（PBoV）.The specific primers and probes were designed according to the VP1 / 2genes of PBoV in GenBank.The result showed that the specificity of this assay was high without any cross-reaction with PPV,PRRSV and PCV.A good linear correlation was demonstrated in the standard curve for the real-time PCR assay within the range of 10^1 to 10^7 copies / mL,with a correlation coefficient of 0.997.The detection limit the real-time PCR assay was 10 1 copies / mL of positive plasmids,indicating agood sensitivity.This real-time PCR based on TaqMan probe technology is a high sensitive and specific for detecting PBoV.

关 键 词：猪博卡病毒 实时荧光定量PCR TAQ Man探针 

分 类 号：S852.659.2[农业科学—基础兽医学]