Cutaneous leishmaniasis (CL), an endemic disease in the littoral zones of the Mediterranean area, the Middle East, East Africa, and especially in Libya, has not been fully documented. The present study clarifies the clinico-epidemiologic profile of CL and the molecular genotyping of the Leishmania spp. in the Nalut district, Libya. Two hundred and twenty-three CL patients were examined at the out-patient clinics of Nalut Hospital from March 2006 to February 2007. CL was diagnosed by clinical, microscopic, culture, polymerase chain reaction (PCR), and PCR-restriction fragment length polymorphism (RFLP) analyses. The disease was observed year-round, with the highest prevalence between November and February. Fifty-nine percent of patients were younger than 20 yr. Nodulo-ulcerative lesions, indurated ulcers, papulo-ulcerative lesions, and subcutaneous nodular lesions were observed in 170, 25, 15, and 13 patients, respectively. Two hundred patients (89.7%) had dry type of lesions, whereas 23 patients (10.3%) presented a wet type of lesion. One hundred and fifty-nine (71.3%) of 223 patients were confirmed positive for CL by the presence of the amastigote form of Leishmania by stained Giemsa smear, and 170 (76.2%) were positive according to the presence of the promastigote form of Leishmania by culture in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS). PCR confirmed 203 (91.0%) positive cases. Genotyping of Leishmania spp. by RFLP analysis revealed that L. tropica was the most common species at all ages, and L. infantum was second under 20 yr of age. In summary, CL is endemic in the Nalut district, Libya; PCR was the most sensitive parasite diagnostic test, and L. tropica was the most common species.