关键词: 伪狂犬病毒, gE基因, 地高辛标记探针, 斑点杂交, 野毒株

Abstract: A 304 bp fragment of gE gene of PRV was amplified from recombinant plasmid pMD18-T-gE and labeled with digoxigenin as DNA Probe. Dot blot hybridization was developed for detection of PRV wild strains. The recombinant plasmid DNA, PRV Bartha-k61DNA, PPV DNA, PCV DNA,PRRSV cDNA and CSFV cDNA were tested by this method. The results showed that positive signal appeared only in recombinant plasmid, while PRV Bartha-k61 DNA, PPV DNA, PCV DNA, PRRSV cDNA and CSFV cDNA were all negative. As little as 4 pg of known positive target DNA (PCR product of gE) could be detected with the digoxigenin-labeled probe. 11 samples collected from cases with reproductive disorders were detected by this probe, and among which, 4 samples were positive. The results of dot blot hybridization were in accordance with that of PCR, it showed that the probe could be used in the clinical diagnosis of pseudorabies.

Key words: pseudorabies virus, gE gene, digoxigenin-labeled probe, dot blot hybridization, wildtype strain