Development of a Multiplex Polymerase Chain Reaction for Differential Diagnosis of Canary Pox Virus

Abstract

Background and Aims: A multiplex transcription-polymerase chain reaction (m-PCR) was developed for direct detection and discrimination between canarypox virus (CPV) and other avian poxvirus (APV). Materials and Methods: Three compatible primer sets were designed for m-PCR amplification of different loci; fpv126, fpv140, and fpv167 located at highly conserved APV genes. Results: Results showed that m-PCR products of the expected sizes were obtained for all of the primer sets when they were tested either alone or in combination with an artificial mixture of positive controls. Based on the better amplification of fpv167 than other loci, the locus primer set was used to examine tissue samples from canaries clinically diagnosed as AVPinfected. Conclusion: All canary samples were positive for CPV by the m-PCR and virus isolation. The results of the present study indicate the m-PCR assay holds potential to be versatile, rapid, and sensitive for detection of CPV and differentiation of the virus from the other APVs.

Molecular Characterization and Phylogenetic Analysis of Avian Pox Virus Isolated From Pet Birds and Commercial Flocks, in Iran

“…The 4b core protein gene (p4b) of APV that encodes the protein with molecular weights of 75.2 kDa is usually chosen for comparative genetic analysis (3,4,5). Also, amplification of the p4b of APV by PCR has often been used as a molecular tool for the detection of APVs (6). PCR in combination with restriction endonuclease enzyme analysis (REA) followed by sequence analysis of the amplified fragments is used for detection, differentiation and molecular characterization of fowl pox virus isolates (7).…”