羊传染性脓疱病毒PCR检测方法的建立与应用

摘要

为快速检测临床羊传染性脓疱病毒(CPDV)感染,根据GenBank中公布的CPDV毒株序列,通过多毒株B2L序列保守区的比对,设计并合成1对特异性引物,利用PCR方法检测该病毒,并进行特异性、敏感性试验及临床检测.结果显示,该反应的最适退火温度为56 ℃,最适引物量为0.6 μL(20 μmol/L),可以检测到1 pg的DNA,并且能鉴别山羊痘病毒、口蹄疫病毒、小反刍兽疫病毒、蓝舌病病毒.结果表明,该方法特异性高、敏感性强,可用于CPDV临床感染的快速检测.%In order to rapidly detect the Contagious pustular dermatitis virus(CPDV)in clinical practice,a PCR method was developed. According to the published gene sequence of virus in GenBank,a pair of specific primers were designed and synthesized by comparing the conserved regions of B2L sequences of multiple strains. Then the CPDV was detected by the developed PCR method,and the specificity,sensitivity and clinical application were tested. The results showed that the optimum annealing temperature was 56 ℃,the optimum amount of primer was 0.6 μL(20 μmol/L), and the detection limit was 1 pg of DNA. In addition,the method could distinguish CPDV and other viruses containing Goat pox virus,Foot and mouth disease virus,Peste des petits ruminants virus and Blue Tongue virus. As a result, a PCR method with high specificity and strong sensitivity was established,and it could be used in rapid detection of CPDV.