A polymerase chain reaction (PCR) was developed to detect a recently described pigeon adenovirus (PiAV). Primers located in the fiber gene of PiAV amplified a PCR fragment solely from PiAV DNA, whereas all 12 serotypes of fowl adenoviruses (FAV1 to FAV12), some of them able to infect pigeons, did not react. A PCR fragment of 967 base pairs was amplified from three different isolates serologically typed as PiAV and from some pigeon liver samples showing morphological and histological signs of an adenovirus infection. Those samples did not react with a published primer pair (H3/H4) able to detect FAV, demonstrating the specificity of both PCRs to react exclusively with the respective pathogen, PiAV or FAV. The presented PCR is a suitable diagnostic tool to gain further insight into the epidemiology of PiAV infections in pigeons.