Detection of LPG2 and GP63 genes sequencing in patients with cutaneous leishmaniasis

Cutaneous Leishmaniasis (CL) is a significant health issue, particularly in Iraq, with a notable increase in cases during the winter months. This cross-sectional study was conducted at Ramadi Teaching Hospital, selected due to the rising incidence of CL. A total of 10 patients, aged 18-40, with an average age of 29, participated in the study, which ran from December 10, 2023, to March 2024. Blood samples were collected from the lesions of CL patients using capillary tubes, ensuring sufficient volume by using multiple tubes per sample. Patients were undergoing treatment with Pentostam as prescribed. The collected blood was transferred to tubes containing sodium chloride solution and stored at- 20 °C until DNA extraction. After storage, genomic DNA was successfully extracted from the samples. This DNA served as a template for polymerase chain reaction (PCR) analysis, utilizing specific primers designed to amplify genes associated with virulence factors. Among the 10 samples, 7 tested positive for DNA extraction, while 3 were negative. Of the 7 positive samples, PCR electrophoresis indicated that 3 were positive for the GP63 gene and 4 were negative. Conversely, 4 samples were positive for the LPG2 gene, while 3 were negative. A control group was included to facilitate comparison during PCR electrophoresis for both genes. Analysis confirmed that Leishmania major is the sole causative agent of CL in Anbar.