Visceral leishmaniosis is an important zoonotic disease in Iran. The agent, vector and reservoir are Leishmania infantum, Phlebetomous major and dogs, respectively. Thus far, no data has been reported about the prevalence of visceral leishmaniosis in human and dog in Torbat-e- Heidareah area and the present study has been performed to determine the frequency of L. infantum infection in stray dogs. One hundred blood and skin samples were collected from 2014 to 2015. The sera samples were examined by Indirect Immunofluorescent Antibody Test (IFAT). The skin samples of seropositive and seronegative in association with skin lesions were examined by PCR method. In this study, antibodies against L. infantum were detectedin 3 (3%) sera samples of dogs and kDNA Leishmania spp. was detected in 4 (4%) of seronegative dogs by PCR. L. infantum was detected in positive samples by semi nested PCR. Based on serological and molecular results, 7% of asymptomatic stray dogs of Torbat-e- Heidareih area were infected with L. infantum and may be acted as reservoirs of visceral leishmaniosis

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