Abstract: A pairs of primers and a Taqman-MGB probes were designed and synthesized according to the the nucleotide sequence of the gE gene of pseudorabies virus(PRV) available in GenBank，and real-time TaqMan-MGB fluorescence quantitative PCR for distinguishing the wild strain and gene-deleted vaccine strain of PRV was established successfully. It was demonstrated that the established TaqMan-MGB quantitative PCR assay could detect 2.23×101 copys?μL-1 of plasmid DNA . Sensitivity and positive rate for clinical sample of TaqMan fluorescent quantitative PCR were higher than routine PCR,and its sensitivity Was 100 times higher than that of the routine PCR.,and had no cross reaction with classical swine fever virus(CSFV)，porcine reproductive and respiratory syndrome virus(PRRSV) ,porcine cireovirus type 2 (PCV2) and Porcine parvovirus (PPV) The 30 suspected material were detected by TaqMan-MGB fluorescence quantitative PCR and routine PCR , respectively, the positive detection rate were 40% and 33%, the coincidence rate was 90% .The real-time TaqMan fluorescence quantitative PCR assay which is specific，sensitive,rapid and accurate can be used for the diagnosis of PRV infection． Key words：pseudorabies virus(PRV)；TaqMan-MGB；real-time PCR