Prevalence and Molecular Identification of Leishmania species in District Mohmand, Khyber Pakhtunkhwa, Pakistan

Abstract

Leishmaniasis is a vector-borne disease that is spread worldwide by the sand fly. It is the second-most neglected and fatal protozoan-born disease, according to the World Health Organization. One of the most prevalent anthroponotic diseases in Pakistan, mostly in Khyber Pakhtunkhwa, is cutaneous leishmaniasis (CL). The aim of this study was to investigate the genetic makeup for identification and frequency distribution of cutaneous leishmaniasis in district Mohmand Khyber Pakhtunkhwa, during 2021–2022. A microscopic method is used to identify amastigote in biopsy reports, skin scrapings, or impressions on smears. The microscopic analysis, however, is insufficient to identify the Leishmania parasite at the species level. Two things, prevalence and parasite identification must be considered while controlling leishmaniasis. For this purpose, the people with CL symptoms who were transferred to the Leishmania diagnostic centers were included in this study. The data collected from 239 CL positive patients was statistically analyzed. The blood smears were taken from Leishmania positive patients for molecular investigation. The genomic DNA from each sample was isolated in order to identify the Leishmania species by the PCR method using ITS1. The phylogenetic tree was constructed by Mega X software using reference sequences retrieved from a gene bank. The distribution of CL cases was represented on map a using Arc-GIS software. The possible correlation between the CL frequency and its risk factors was statistically analyzed by the chi-square test. A structured questionnaire containing some demographic questions was filled out by each positive patient. A total of 239 microscopy positive patients were reconfirmed by molecular technique using ITS1 gene, confirmed 216 CL patients for L. tropica species. The male to female ratio recorded was 60.7%) and 39.1% (P = 0.001). The early ages (1-20 years) are found to be more susceptible to CL. The highest number of lesions was found in the facial area (56.9%), followed by the hands (21.8%) and legs (21.3%). In (77.4%) of patients’ single lesion was found whereas the rest had more than one lesion. The lesion's duration ranges from 1-4 months were (68.6%), 5-8 months were (25.5%), 9-12 months were recorded (4.2%), and more than 12 months were (1.7%) respectively. The current research showed that anthroponotic cutaneous leishmaniasis (ACL) was observed in the district Mohmand in Khyber Pakhtunkhwa. DRSML QAU xii Cutaneous leishmaniasis is still a major public health problem in this region. Cutaneous leishmaniasis was observed more frequently during the winter season. It will require more efficient measures of control in the future to prevent and eradicate the infection.