Canine leishmaniosis (CanL) caused by Leishmania spp. is a zoonotic protozoon disease. It is widespread in most parts of the world including the Mediterranean basin. The present study was carried out in order to determine the prevalence of CanL in dogs in Kayseri and vicinity by nested-PCR. A total of 300 asymptomatic dogs were sampled randomly. Blood samples taken from the vena cephalica antebrachii were collected into tubes containing EDTA. Furthermore, lymph samples were taken from 14 dogs while bone marrow, spleen and liver biopsies were taken from three dogs. The DNA's obtained from these samples were examined for the presence of Leishmania spp. by nested-PCR which amplified the small subunit ribosomal RNA (ssr RNA) gene. According to the results of the nested-PCR, none of the 300 dogs were Leishmania spp. DNA positive.