摘要： 根据猪伪狂犬病病毒(PRV)gE、gH基因序列,设计合成了2对特异性引物,分别建立gE和gH基因的单项PCR扩增方法。通过对PCR扩增条件的优化,建立了鉴别PRV强毒和疫苗毒的双重PCR检测方法,即利用一次PCR反应,从强毒株基因组中可同时扩增出2条大小分别为429 bp(gE基因)、355 bp(gH基因)的特异性片段,从弱毒株DNA中仅扩增出1条大小为355 bp的片段,而猪圆环病毒2型和猪细小病毒DNA扩增结果均为阴性。敏感性试验表明,所建立的双重PCR检测方法最低有效检测量为1×10²TCID₅₀/0.1 mL。该方法适合对PRV强毒和疫苗毒的快速鉴别诊断。

关键词: 伪狂犬病毒, 双重PCR, 检测, 强毒株, 疫苗株

Abstract: Based on the gE and gH genes of pseudorabies virus (PRV), two pairs of special primers were de- signed, respectively, single PCR assays for gE and gH genes were established. Under the optimized amplification conditions, the duplex PCR method was established to differentiate virulent and vaccine strains of pseudorabies vi- rus. In a duplex PCR assay, two fragments of 429 bp for gE gene and 355 bp for gH gene were simultaneously am- plified from pseudorabies virus virulent strains DNA, only a 355 bp fragment for gH gene was amplified from pseud- orabies virus vaccine strains DNA, whereas no PCR products were amplified from porcine circovirus type 2 and por- cine parvovirus. Then the detection limit of duplex PCR was estimated to be 1 × 10²TCID₅₀ /0. 1 mL. This method is suitable for differentiation of wild-type pseudorabies virus and vaccine strains in clinical specimen.

Key words: Pseudorabies virus, Duplex PCR, Detection, Virulent strains, Vaccine strains

中图分类号: 

• S828