Development of real-time TaqMan-quantitative RT-PCR assay for detection of porcine reproductive and respiratory syndrome virus

The probes and primers were designed and synthesized according to the conserved gene ORF7 of PRRSV available in GenBank,and then reaction parameters were optimized to develop a real-time TaqMan-quantitative RT-PCR assay.The serum and other tissue samples from pigs on farms in Dongguan,Zengcheng,Zhanjiang and other areas of Guangdong Province were detected by using the established quantitative RT-PCR assay,and the results was compared with that of routine RT-PCR.The developed quantitative RT-PCR assay could detect 5.0×10~(0) copy/μL of plasmid DNA and its sensitivity was 100 times higher than that of the routine RT-PCR,while the results of the quantitative RT-PCR were the same as that of the routine RT-PCR.Three samples were examined using the real-time RT-PCR repeatedly and the results indicated that the real-time quantitative RT-PCR was reproducible and could be used for the(diagnosis) of PRRSV infection.