区分猪伪狂犬病病毒gE基因缺失疫苗株和野毒株的gE-PCR方法的建立

A gE-PCR for Differentiation Pseudorabies virus gE-Vaccine Strain from Wild-type Strains

根据编码猪伪狂犬病病毒gE基因保守序列,设计合成一对引物,通过优化PCR反应条件,成功地从猪伪狂犬病病毒感染的细胞中扩增出预期的178bp片段,而猪繁殖与呼吸综合征病毒、猪细小病毒、猪瘟病毒、猪流感病毒和猪伪狂犬病病毒gE基因缺失株均未扩增出相应的片段,经PCR扩增产物测序鉴定,证实了该扩增片段为预期目的片段;敏感性试验表明,该体系可检测到102TCID50的猪伪狂犬病病毒。本方法的建立能够区分基因缺失疫苗免疫猪和野毒感染猪,使猪伪狂犬病病毒的检测更为快速、准确。

A PCR detection method was developed using a pair of primers designed from a conserved region of the pseudorabies virus（PRV）gE gene. This method used all optimized PCR eonditions to detect the presence of PRV sequence. A 178 bp fragment was amplified from PRV infected PK-15 cells, but not from PRRSV,PPV, HCV,SIV and PRV gE strains. This amplified fragment is specific for PRV DNA based on DNA sequencing. The sensitivity is 102TCID50. The result showed that this PCR method could differentiate PRV gE- vaccinated swine from wild-virus infected swine. It could he a fast and specific detection method for PRV.