Leishmaniasis is a multisystem disease, and have a broad spectrum ranging from skin lesions to systemic disease. Therefore diagnosis must be supported with laboratory results. We analyzed 153 smears, aspiration, blood and bone narrow samples collected from patients suspected with cutaneous and visceral leishmaniasis. The specificity and sensitivity of the four methods (culture, smear, miniexon-PCR-RFLP and ITS1-PCR-RFLP) were detected and Leishmania species were determined. The ITS1-PCR-RFLP method was found that the highest sensitivity and specifity. L. infantum and L. tropica were identified by molecular methods from samples. As a result, ITS-1-PCR has a high sensitivity and specificity and easily applicable method. However, it requires the miniexon-PCR or ITS1 sequencing the discrimination of the L. donovani complex. L. infantum is a agent both visceral and cutaneous leishmaniasis in our region.