The role of papillomaviruses (PVs) in the development of canine cancers is controversial. However, recently a novel canine PV (CPV3) was detected in a dog affected with a condition reminiscent of epidermodysplasia verruciformis (EV). The aim of the present study was to investigate the seroprevalence of CPV3 by using generic enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against either canine oral PV (COPV) or CPV3. Therefore, the capsid proteins of both PV types were expressed as glutathione S-transferase fusion protein antigens and adsorbed to glutathione-casein-coated ELISA plates. After showing that PV type-specific antibodies could be detected in the sera from dogs with confirmed COPV or CPV3 infection, CPV3-and COPV-seropositive samples were detected in two sets of canine sera collected in Switzerland and South Africa, respectively. We found specific antibodies against COPV and CPV3 among the tested sera and also a large number that were positive for both antigens. The seroprevalences of PV antibodies of 21.9% (COPV) and 26.9% (CPV3) among the tested dogs from South Africa were higher than those among the dogs from Switzerland at 10.5% (COPV) and 1.3% (CPV3). Our data suggest a need for further CPV-related seroepidemiological surveys in different countries, especially in the context of clinical manifestations and possible breed predispositions. For this purpose, the newly developed ELISAs can be a useful tool.Papillomaviruses (PVs) are small, icosahedral, nonenveloped DNA viruses with mucosal and skin tropism which are associated with various hyperplastic, dysplastic, and neoplastic conditions in humans and animals (21). More than 100 types of human PVs (HPVs) have been described, and more are continuously being discovered (8). The role of HPVs in the development of human diseases, i.e., cervical carcinomas or epidermodysplasia verruciformis (EV)-associated skin carcinomas, is well established, and pathomechanisms have been the focus of many studies (14,23,30,35,48). Indeed, both PCR-and enzyme-linked immunosorbent assay (ELISA)-based techniques are commonly used to carry out HPV-related epidemiologic studies (2, 25).In contrast, the role of PVs in the development of canine cancers remains uncertain, although a clinical counterpart to human EV has been described in dogs with PV infections (28). Until now, only four canine PVs (CPVs) have been unambiguously identified and sequenced (7,42,43,46). The first and best-known CPV, canine oral PV (COPV), affects mostly juvenile dogs and causes warts with a typical cauliflower-like appearance located in the oral cavity and other mucous membranes (32). Since COPV-affected dogs recover spontaneously and specific antibodies protect them from reinfection, COPV infection is not considered a life-threatening condition (32). Although COPV infection is endemic in most dog populations, there never was a great demand for serological surveys, and systematic serological data concerning the prevalence of COPV infection are thus far unavailable. A dis...