Molecular profiling and risk factors assessment of cutaneous leishmaniasis in an expanding endemic focus of Punjab, Pakistan

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Abstract
Cutaneous Leishmaniasis (CL) is a vector-borne disease caused by Leishmania species, which is responsible for anthroponotic and zoonotic CL in Pakistan. Due to the unavailability of reports on the molecular epidemiology and risk factors assessment of CL in Khushab, Punjab, Pakistan, this study aimed to investigate the incidence rate of CL, associated risk factors, and molecular identification of Leishmania species in this region. The clinical and demographic data of 423 suspected CL patients were collected. Their skin scrapings and blood samples were analyzed microscopically for amastigote detection and for genomic DNA extraction, respectively. A partial fragment (~ 400 bp) of ITS1 gene was amplified followed by restriction fragment length polymorphism (RFLP) assay and sequencing for species identification. Meteorological data were obtained from the Pakistan Meteorological Department, Lahore. Statistical analysis was conducted using Univariate and multivariate regression models using SPSS. Microscopy confirmed 41.8% of the cases, while PCR detected Leishmania DNA in 33.8% of the samples. CL infection was more common in females (57%) and in the < 1–20 age group (54.8%). The majority of the CL lesions were dry (63.2%), single (69.4%), predominantly affecting the facial region (46.3%), occurred with a duration of less than 2 months (75.7%), and were less than 1 cm in size (62.7%). Cases were highest in January (19.3%) and lowest in June (2.6%). The average temperature (22–27 °C) and reduced rainfall (4 mm) were key factors for sand-fly breeding. The infection rate was high (44%) among the residents of Padhrar. Molecular analysis (ITS1-RFLP) confirmed the presence of L. tropica, with ITS1 sequences exhibiting 100% similarity to isolates reported from India, Pakistan, Iran and Afghanistan. This study highlights the increasing number of CL cases in non-endemic areas, underscoring the importance of public health education, improved disease control programs, and cross-sector collaboration to mitigate the impact of CL and prevent its further spread.
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  • Abdul Rahman Osmani

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