Molecular Typing and Phylogenetic Analysis of Cutaneous Leishmania from Iraqi Patients

Abstract

This study aimed for molecular characterization of local isolates of L. tropica and L. major and their
relatedness to reference isolated from neighboring countries. Exudates and dermal tissues were collected
from the cutaneous lesions of 50 newly diagnosed patients with CL. Leishmania DNA was extracted from
lesion exudate and 5.8S rRNA gene was amplified using two sets of specific primers through nested PCR. A
total of 12 PCR products were directly sequenced and the resultant sequences were aligned with reference
sequences in National Center for Biotechnology Information (NCBI). A phylogenetic tree was constructed
for local Leishmania isolates with the reference isolates using Mega 6 software. According to alignment
results out of 12 sequences, 9 sequences were belonging to L. major, while 3 sequences were belonging to
L. tropica. All local L. major isolates were very close to reference isolate KP773404.1 (an Iranian isolate),
while all local L. tropica isolates were very close to reference isolate KY612611.1 (an Iranian isolate) with
an identity of 98%. Phylogenetic trees confirmed that closest isolates for local Leishmania (whether L.
tropica or L. major) was the Iranian isolate. These data indicate that local isolates of L. tropica and L. major
are closely related to Iranian isolates.