Molecular identification and spatial distribution of cutaneous leishmaniasis cases in Markazi Province, central Iran, using ITS1 PCR RFLP and GIS

Cutaneous leishmaniasis (CL) remains a considerable public health concern in Iran. This study aimed to characterize Leishmania spp. among available diagnostic specimens and describe the spatial distribution of cases reported in the Markazi Province during 2021–2022. In this cross-sectional study, all 210 microscopically confirmed CL cases were analyzed using a total census sampling method. A subset of 42 cases with available diagnostic smears was evaluated by internal transcribed spacer 1-polymerase chain reaction-restriction fragment length polymorphism (ITS1-PCR-RFLP) for species identification, followed by sequencing of four representative amplicons. Spatial distribution of the 42 molecularly characterized cases was mapped using a Geographic Information System (GIS; ArcGIS 10) based on recorded coordinates. Among all 210 cases, CL was more frequent in males (57.1%) with the highest proportion in the 40–49-year age group (20.5%). Lesions were predominantly ulcerative with exudate and localized to the extremities. All 42 molecularly analyzed samples were identified as Leishmania major , though this convenient subset may not fully represent the overall species composition across the entire province. Phylogenetic analysis confirmed that the sequenced isolates clustered within the L. major clade. GIS-based descriptive mapping indicated that analyzed cases were localized primarily in Saveh and Mahallat counties. These findings provide an updated baseline for CL in Markazi Province; however, systematic molecular surveys and vector-reservoir investigations are needed to fully delineate species distribution and transmission dynamics across all counties.