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.