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The present study was performed to identify the species and strain of Leishmenia parasite isolated from different endemic areas. It was carried out on cutaneous leishmeniasis (CL) in Mid-Eufrate Provinces, Iraq. A total of 126 cases comprising 48(38%) females and 78(62%) males were studied. The highest infected age group was 21-30 years with a rate of 27.8% and the lowest rate was 11.9% at the age group of 10 and less years. Geographical distribution of CL among hospital patients indicated that rural areas were with highest rate (54%) while in urban areas were with (46%). The highest rate of multiple lesions was 73% in comparison with single lesion (27%). The disease was in wet type (82.5%) while (17.5%) in dry type. Direct smears gave positive result in 90 (71%) patients under light microscope. Out of 126 cases, 83(65%) gave positive growth. Out of 83 postive growth culture, only 52 specimens were studied by nested - PCR. It was found that 45(86.5%) cases in the generation of a 560 bp DNA and 7(13.4%) patients displayed a fragment of 750 bp, corresponding to L. major and L. tropica, respectively. Cellulose acetate electrophoresis (CAE) was performed for 20 mass cultivated culture.