Real-time PCR protocol for the detection of porcine parvovirus in field samples

This report describes a real-time polymerase chain reaction assay with SYBR® Green for detection of a broad range of porcine parvoviruses (PPV) and accurate virus quantification in porcine tissues. The assay targets the VP2 gene of PPV and the porcine genomic c-myc gene for normalization. The detection limit of the SYBR® Green reaction was shown to be equivalent to 6 × 100 to 6 × 101 PPV copies/reaction and the overall detection limit equivalent to 0.1 TCID50/100 μl. The assay was linear over a 107 dilution range of template concentrations. Other porcine pathogens involved in reproductive disorders such as porcine circovirus 2 (PCV-2), porcine reproductive and respiratory virus (PRRSV), Aujeszky's disease virus (PRV) and other parvoviruses such as feline parvovirus (FPV), canine parvovirus (CPV), minute virus of canines (MVC) and a human parvovirus (B19) were not detected by this assay.