摘要
目的建立猪伪狂犬病病毒(PrV)的PCR检测方法。方法根据PrV gB基因序列,应用primer 5.0软件自行设计、合成一对引物进行PCR反应,优化反应条件,建立检测PrV的PCR方法,并应用于临床样品的检测。结果以PrV上海株细胞培养物DNA为模板,扩增出263 bp的特异性条带,对扩增产物进行克隆测序和BLAST在线比对,与GenBank中PrV的gB序列一致。对临床样品进行PCR检测,与病毒分离结果一致。结论所建立的PCR方法敏感、特异,可用于实验用猪伪狂犬病病毒的快速检测。
Objective To establish the method ofpolymerase chain reaction （PCR） for detection of the pseudorabies virus （PrV） in pigs. Methods A pair of primers were designed and synthesized according to the sequence of gB gene of PrV. PCR method for PrV was established by optimizing the reaction parameters, and utilized for detection of PrV in clinical samples. Result The 263 bp product was amplified specifically by using the DNA of PrV SH strain as template. The amplified product was cloned into pMD18-T vector and sequenced. The identity of nucleotide sequence of this fragment and gB gene of genbank was 99.2%. The identical results were obtained by PCR and virus isolation of clinical samples. Conclusion The PCR method established is sensitive and specific for the rapid detection of PrV in pigs.
出处
《实验动物与比较医学》
CAS
2008年第2期114-117,共4页
Laboratory Animal and Comparative Medicine
基金
上海市科技发展基金(024909006)
上海市农科院青年基金(2004-02)资助
关键词
伪狂犬病病毒
PCR
检测
Pseudorabies virus （PrV）
Polymerase chain reaction （PCR）
Detection
