Isolation and molecular detection of pigeon pox virus in Assiut and New Valley governorates

Abstract

Pigeon Poxvirus (PPV) was detected in eight pigeons suffering from wart like nodular lesions in two Egyptian governorates (Assiut and New Valley) during summer 2018. Different serological and molecular techniques were carried out for isolation and detection of the virus on chorio-allantoic membranes (CAM) of specific-pathogen-free (SPF) embryonated chicken eggs. The characteristic pock lesions were detected on CAMs, whereas PPV was isolated. Electron microscopy revealed enveloped brick shaped Avipoxvirions. The neutralizing antibodies against PPV were detected in six out of eight samples. Serum neutralization test revealed a neutralization index of ≥ 1.6, while ELISA revealed an S/P ratio of ≥ 1.4 in the affected pigeons. Nucleotide sequence of P4b of Pigeon poxvirus isolated from nodule 1 sample (PPVNV1), revealed 100 % nucleotide identity to PPV and only 90 % nucleotide identity with Fowl poxvirus (FPV). P4b locus based SYBR green QPCR produced PPV amplicons of 77.33–77.83 °C melting temperature (Tm). QPCR SYBR green assay successfully differentiated PPV from FPV amplicon which revealed a dissociation curve of Tm =75.85 °C. This is the first report discriminating PPV from FPV based on SYBR green qPCR approach of P4b locus. This isolated local Egyptian strain can be used in vaccine production for optimal vaccination strategy.