Development and preliminary application of TaqMan fluorescence quantitative RT-PCR assay for detection of porcine Sapovirus

The primers and probes were designed and synthesized according to the conserved VP1 sequences of porcine Sapovirus (SaV), and a TaqMan fluorescence quantitative RT-PCR assay were developed by optimizing the reaction conditions. Results showed that the fluorescence quantitative RT-PCR assay could detect 16.1 copies·μL-1of plasmid DNA, while the sensitivity of the routine RT-PCR was 1.61 × 103copies·μL-1. 216 stool samples were then detected by the established quantitative RT-PCR assay, and the results were compared with that of routine RT-PCR. It also showed that the sensitivity of established method was higher than that of the routine RT-PCR. Phylogenetic analysis indicated that all of the 4 SaV strains we had identified belonged to G?, and shared 100% nucleotide homology with another Shanghai porcine SaV strain (FJ387164). The TaqMan fluorescence quantitative PCR assay, which is more specific, sensitive and accurate, can be used for the epidemiological investigation and diagnosis of porcine SaV infection.