Cutaneous leishmaniasis (CL) is a zoonotic disease with 1 to 1.5 million annual incidences. Microscopic examination of the Giemsa stained slides is the most common diagnostic method for CL. However, this method cannot distinguish leishmania species. Hence the present study was conducted to identify leishmania species by high-resolution melting (HRM) analysis in the newly emerged foci of CL in Sabzevar, northeast of Iran. In this cross-sectional study, fifty patients with suspicious cutaneous lesions referring to the designated health center for diagnosis and treatment of CL in Sabzevar during 2017–2018, were recruited. All collected samples and prepared slides were stained for microscopic examination and then undergone HRM real-time PCR (HRM-PCR) assay to identify species of Leishmania parasites. The results of HRM-PCR technique showed that Leishmania major (L. major) was the dominant causative parasite in the newly emerged foci whereas L. tropica (L. tropica) was positive only in two patients. This was the first time that 7SL RNA-HRM-PCR assay was performed to precisely identify leishmania parasites in the northeast of Iran. We proved the newfound foci in which both L. major and L. tropica were present. In contrast to the recent studies which identified only L. major in the region, we showed that L. tropica was still present.
