【摘要】 目的建立犬细小病毒的核酸诊断方法(PCR方法)并应用于临床诊断。方法根据犬细小病毒VP2基因序列设计引物,以犬细小病毒标准株、犬传染性肝炎病毒和正常增殖细胞DNA为模板,犬细小病毒扩增出221bp的特异带,将PCR产物连接到pMD18_T Vecter,转化大肠杆菌JM109菌株。重组质粒经测序并经blast比较,与GenBank中已登录的犬细小病毒VP2基因序列进行比较。同时将建立的PCR方法应用于30份犬粪便和病犬组织的检测,并测序。结果PCR产物测得的序列与GenBank的基因序列同源性为100%,粪便检测均无犬细小病毒的特异带,从病犬组织中有6份样品扩增出221bp的特异带,经测序比较,同源率为100%。结论建立了犬细小病毒的PCR检测方法,并能应用于临床诊断。更多还原

【Abstract】 Objective To establish a polymerase chain reaction method for canine parvovirus dection and to utilize the PCR for diagnosis of parvovirus in canine samples.Methods PCR primers were designed according to the VP2 Gene of CPV.221bp products was obtained with the template from CPV strain,while no band was found in infectious canine hepatitis virus.This amplified product was cloned into pMD18_T vector and transferred into E.coli JM109.The recombinant plasmid was sequenced.30 feces and 12 tissue samples were detected of CPV by PCR method.Results The identity of DNA sequence of this fragment between CPV strain and gi54646342/gi39748709/gi57903634 was 100%.All of 30 feces samples was negative,221bp fragment was amplified from 6 of 12 tissue samples.Conclusion A PCR method is established for the detection of parvovirus.The PCR assay is an sensitive and specific technique for parvovirus diagnosis.更多还原