Introduction & Objective: Leishmaniasis is one of the most important infectious diseases in the world. This disease is reported from more than 15 provinces of Iran as a health problem. Identification of reservoirs and characterization of parasite species in patients is very important for identifying the epidemiological aspects of disease and planning for control program.

Materials & Methods: This survey was performed as a descriptive study by PCR-RFLP method in Damghan district. Study was carried out on giemsa-stained smears from cases of leishmaniasis and rodents which collected in infected villages that leishmanal infection was confirmed by microscopic observations. DNA of giemsa-stained slides was extracted and followed by conventional PCR technique using specific primers of ITS1. PCR products were digested with a restriction enzyme (HaeIII).

Results: Totally 25 smears from human skin lesions and 8 rodent ear scrapings were examined by PCR. Electrophoresis results showed that leishmania major was present in patients and rodents giemsa-stained slides.

Conclusion: PCR-RFLP is an effective method to identify leishmania species from giemsa-stained smears which have been collected from humans and animal reservoirs. Using this method identification of causative species of leishmaniasis is possible without sequencing.