INTRODUCTION: Cutaneous Leishmaniasis (CL) is a vector-borne disease caused by a protozoan parasite and considered a public health challenge in many countries, including Iran. Recent research has focused on the role of Leishmania RNA virus (LRV) in the pathogenesis of cutaneous and mucosal forms of leishmaniasis. This study assessed the presence of LRV2 and its genotype diversity among isolates in Ilam province, western Iran, an old focus of Zoonotic CL. MATERIAL AND METHOD: Thirty-two lesion specimens were collected from CL patients and subjected to PCR analysis using species-specific primers for Leishmania identification and detection of LRV2. The amplified products were sequenced, and phylogenetic analysis was performed. RESULT: All samples were identified as L. major. The PCR showed that 4 of 32 (12.5%) clinical cultures were LRV2 positive. Phylogenetic analysis revealed high nucleotide sequence identity (96-99%) with LRV2 isolated from L. major strains from Iran, Turkey, and Uzbekistan. The four sequences analyzed revealed four distinct haplotypes, with high genetic variability among the isolates. CONCLUSION: This study provides the first identification and phylogenetic analysis of LRV2 in CL patients from western Iran and highlights the extensive genetic diversity among LRV2 isolates across different regions, which may have significant implications for disease pathogenesis, transmission patterns, and clinical manifestations.