The island of Cyprus is located in the Eastern Mediterranean, where leishmaniasis is endemic. Although human visceral and cutaneous leishmaniasis (VL and CL) cases have already been documented on the island, there are limited data on the Leishmania species in northern Cyprus. In this report, we present a CL case diagnosed by both microscopic examination and quantitative real-time PCR (qPCR). The patient, a 79-year-old man residing in northern Cyprus, developed an ulcerative lesion on his left leg. The lesion was surgically excised for histopathological examination, and tissue sections were stained with hematoxylin and eosin (H&E). Microscopic examination of H&E-stained tissue sections revealed Leishmania amastigotes. To confirm the diagnosis and identify Leishmania species at the molecular level, DNA was extracted from the paraffin-embedded tissue sections. Following deparaffinization, qPCR targeting the Leishmania-specific Internal Transcribed Spacer 1 region (located between SSU and 5.8S rRNA genes) was performed. In the qPCR assay, the infecting agent was identified as a member of the L. donovani complex, presumptively L. infantum, based on the melting curve analysis. Our findings provide molecular evidence for the presence of leishmaniasis in northern Cyprus and contribute to addressing the lack of molecular data in the region. Our study also suggests that, due to the zoonotic nature of the identified pathogen, continuous vector and reservoir control programs should be implemented in the region to prevent the spread of the disease.