Establishment of a real-time fluorescent quantitative RT-PCR assay for detection of porcine PD-1, PD-L1 and PD-L2 genes.

To establish a real-time fluorescent quantitative RT-PCR assay targating on porcine PD-1,PD-L1 and PD-L2 genes,specific primers and fluorescent probes were designed accrodingly.The reaction conditions for the real-time fluorescent quantitative RT-PCR were optimized based on the serially-diluted recombinant plasmids pMD-PD-1,pMD-PD-L1 and pMD-PD-L2.The assay worked well with a good coefficient correlation(r20.99) and amplification efficiency(96%) while the template concentration was from 1×102 to 1×108 copies/μL.The sensitivity was sufficient to detect at least 100 copies of the samples.The established assay was able to detect the expression of the three genes in porcine peripheral blood mononuclear cell,indicating that the assay was highly sensitive,specific and well reproducible,and suitable for detection of clinical samples.