犬细小病毒病PCR检测方法的建立及应用

Establishment and Application of PCR Detection Methods of Canine Parvovirus

犬细小病毒病感染是由犬细小病毒（cPv）引起犬只死亡的最重要传染病之一，目前诊断该病的方法主要是胶体金快速检测试板法，虽然检测方法较简便快速，但其敏感性较差。通过对NCBI网站GenBank发表的CPV-2基因组序列的分析，选择该病毒VP2基因保守序列设计引物，通过对阳性病料扩增及扩增产物测序，与NCBI发表的相关基因对比，同源性在99．8％-100％，成功建立了特异性、灵敏度高的PCR方法，最低只需2．5PgDNA模板。在对28例可疑CPV病例检测中，本方法检出25例阳性、3例阴性，阳性率为89．28％；胶体金检测板检测出20例阳性、8例阴性，阳性率为71．42％，证实PCR方法比胶体金检测更敏感。

Canine parvovirus infection is caused by canine parvovirus （CPV）, and is one of the most important infectious diseases causing dog deaths. The current diagnosis of the disease mainly uses the colloidal gold rapid detection test plate method. Although this detection method is much simple and rapid, but its sensitivity is poor. Based on the analysis of CPV-2 genome sequence published in the website NCBI GenBank, the author selected the gene conserved sequence of the virus VP2, then designed primers to amplify the material through positive diseases and sequencing of PCR products, and compared them with related genes published in NCBI. We found they were homologous from 99.8 % to 100%, and successfully established the PCR method with specificity and high sensitivity, and the minimum was just 2.5 pg DNA template. In 28 cases of suspicious CPV case detection, this method detected 25 cases as positive, and 3 as negative, with 89.28% positive rate; the colloidal gold assay plates detected 20 cases as positive, and 8 as negative, with 71.42% positive rate. This confirmed that the PCR method was more sensitive than the colloidal gold assay test.