Previous serological surveys have reported the presence of different organisms in cats from Spain but little reports exist about the exact identity of these organisms. The purpose of the study reported here was to assess the presence of DNA of several vector-borne infections in a population of cats from Barcelona area. One hundred blood samples obtained from cats admitted to the UAB-VTH were entered into the study and classified as healthy (n = 48) or unhealthy (n = 52). EDTA-blood samples were assayed for Leishmania infantum, Ehrlichia spp., Anaplasma spp., Rickettsia spp., Bartonella spp., Hepatozoon spp., Babesia spp. and Theileria spp. DNA by means of PCR amplification and amplicons obtained were sequenced. Prevalence of infectious agents found were Leishmania infantum (3%), Ehrlichia/Anaplasma sp. (1%), Hepatozoon felis (4%) and Bartonella clarridgeiae (1%). Cats being less than 5 years old had more probability of having at less one PCR positive result (P = 0.028). The results of this study show a low prevalence of several vector-borne pathogens among cats from Barcelona area. Although higher feline seroprevalences are previously reported, they evidenced exposure and probably overestimate the real or active degree of infection. However, it is important to maintain a high index of suspicion on these infectious diseases, both in sick and asymptomatic cats, and molecular techniques could aid in the identification of these pathogens. © 2007 Elsevier B.V. All rights reserved.