Rapid detection of pigeon herpesvirus, fowl adenovirus and pigeon circovirus in young racing pigeons by multiplex PCR

Infections of young racing pigeons with pigeon herpesvirus (PiHV), fowl adenovirus (FAdV) and pigeon circovirus (PiCV) are reported frequently. The role of these viruses in the pathogenesis of a disease complex called young pigeon disease syndrome (YPDS) is generally accepted. All of these viruses cause inclusion bodies in the liver so liver samples are particularly useful for the detection of infection. Consequently a multiplex polymerase chain reaction (PCR) was developed for the detection of PiHV, FAdV and PiCV in liver samples from racing pigeons. The detection limits were 101 genome equivalents for the detection of PiHV and PiCV and 103 genome equivalents for FAdV. The absence of PCR inhibitors was shown by the detection of cytochrome B gene as an internal control. No PCR products were amplified from related herpes and circoviruses or negative controls, demonstrating the specificity of the multiplex PCR. The addition of cellular DNA from liver samples or Q-solution to the reaction mix had no influence on its sensitivity. The usefulness of the multiplex PCR was demonstrated by re-investigation of liver samples from young racing pigeons previously tested positive by uniplex PCRs.