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 (R) Green for detection of abroad 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 (R) Green reaction was shown to be equivalent to 6 x 10(0) to 6 x 10(1) PPV copies/reaction and the overall detection limit equivalent to 0.1 TCID50/100 mu l. The assay was linear over a 10(7) 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 (13 19) were not detected by this assay. (c) 2006 Elsevier B.V. All rights reserved.