A duplex PCR assay for simultaneous detection and differentiation of Capripoxvirus and Orf virus

A duplex polymerase chain reaction (PCR) was developed and optimized for simultaneous detection and differentiation of Capripoxvirus (CPV) (including goat pox virus and sheep pox virus) and orf virus (ORFV). Two sets of specific oligonucleotide primers were designed and used for CPV and ORFV, respectively. The duplex PCR DNA products, which consisted of fragments of 413 bp for A29L gene of CPV, and 708 bp for P55 gene of ORFV, were visualized by gel electrophoresis. The developed assay was found to be highly specific and sensitive with a detection limit of 1 plaque forming unit (pfu) for both CPV and CPV.