山羊痘病毒PCR检测方法的建立

Development of PCR Method for Detection of Goat Pox Virus

为了县级兽医实验室能够准确检测山羊痘病毒(Goat pox virus,GTPV),根据GenBank上的山羊痘病毒(GTPV)的基因序列,选取p32基因内的保守序列作为检测目的片段,设计、合成一对扩增引物,特异性扩增该目的片段,扩增产物大小为218 bp,建立了山羊痘病毒PCR检测方法。对所建立的PCR反应体系的灵敏性进行了评价,并用此方法对30份羊口鼻棉试子样品进行了检测。结果显示,该方法最低检测浓度为1.82×10^(5)拷贝/μL。检测样品均为GTPV阴性。本文建立了山羊痘病毒PCR检测方法,具有较高敏感性,可储备为本实验羊痘病毒病原检测方法。

In order to accurately detect goat pox virus(GTPV) in veterinary laboratory at county level,according to the gene sequence of GTPV published in GenBank, the conserved sequence of P32 gene was selected as the target fragment for detection, and a pair of amplification primers were designed and synthesized to specifically amplify the target fragment. The length of amplified products was 218 bp, and a PCR method for detection of GTPV was established. Then the sensitivity of the established PCR method was evaluated, and 30 sheep samples(cotton swabs of nose and mouth) were detected by the method. Results showed that the detection limit was 1.82×10^(5)copies/μL, and all samples were GTPV negative. In this study, a PCR method for detection of GTPV was established, which was highly sensitive and could be reserved for GTPV detection.