This disclosure relates to compositions and methods of their use in detection and identification of a chordopoxvirus in a sample, such as diagnosis of an infection in a subject. The compositions and methods allow for detection and identification of all non-avian low-GC content chordopoxviruses, identification of most known high-GC content chordopoxvirus, and species-specific detection of Canarypox virus, Fowlpox virus, and Sealpox virus.