Development and application of real-time fluorescent quantitative PCR assay to detect pseudorabies virus

A pair of primers and a TaqMan probe were designed according to gE gene nucleotide sequence(EF552427) of PRV in GenBank.The expected fragment was amplified from DNA of PRV-infected PK-15 cells,The PCR product was cloned into pGEM-T Easy vector and sequenced.The positive recombination plasmid was used as a positive quantitative template to establish a standard curve.By optimizing the probe's concentration,Mg2+concentration,primers concentration and the annealing temperature,rea1-time fluorescent quantitative PCR was established.Clinical detection of PRV by rea1-time fluorescent quantitative PCR showed that rea1-time fluorescent quantitative PCR paved the way for the early and rapid detection of PRV and quantitative analysis for the infect degree of PRV.