Molecular Typing of Two Suspected Cutaneous Leishmaniasis Isolates in Baghdad

Duha Bayram,Hayder Zuhair Ali

doi:10.21123/bsj.2021.18.1.0029

Abstract

Leishmaniasis is a group of parasitic diseases caused by Leishmania spp., an endemic infectious agent in developing countries, including Iraq. Diagnosis of cutaneous lesion by stained smears, serology or histopathology are inaccurate and unable to detect the species of Leishmania. Here, two molecular typing methods were examined to identify the promastigotes of suspected cutaneous leishmaniasis samples, on a species level. The first was species-specific B6-PCR and the second was ITS1-PCR followed by restriction fragment length polymorphism (RFLP) using restriction enzyme HaeIII. DNA was extracted from in vitro promastigote culture followed by amplification of kDNA by B6 or amplification and digestion of LITSR/L5.8S. PCR produced bands of ~359 bp and ~450 bp for B6 and ITS1, respectively. Digestion of ITS1 by RFLP revealed two distinct bands of ~150 bp and ~300 bp size. The results reviled that the two isolates belong to cutaneous Leishmaniasis, specifically Leishmania tropica. In conclusion, the confirmation of the studied methods will improve rapid and accurate diagnosis of Leishmania species of the most prevalent Iraqi strain of cutaneous leishmaniasis, L. tropica.

Highlights

Leishmaniasis is one of the neglected diseases caused by infection with protozoan parasites belong to a member of Leishmania species [1]
Cutaneous leishmaniasis is caused by numerous spescies of Leishmnaia and are able to cause human leishmaniasis counting at least twentyone species and subspecies [2]
The samples were previously isolated from skin ulcers of two patients attended ALKarama Teaching hospitals in Baghdad and confirmed as cutaneous leishmaniasis by clinical presentation according to the dermatologist

Summary

Introduction

Leishmaniasis is one of the neglected diseases caused by infection with protozoan parasites belong to a member of Leishmania species [1]. The diagnosis of cutaneous leishmaniasis, in Baghdad hospitals and suburban endemic areas generally relies on clinical presentation, microscopic examination in stained smears, rapid agglutination test and parasite culture due to the lack of developed diagnostic tools, which in most cases, cannot identify the causative species [9,10,11]. Local clinical samples were investigated by two species-specific diagnostic assays including B6-PCR amplification and ITS1PCR-RFLP for cutaneous leishmaniasis molecular identification of parasite species as described by Altamemy [21] Kermanjani et al [22] and Schönian et al [23]. The studied methods demonstrated rapid and sensitive assays for typing of Iraqi isolates of Leishmania on a species level

Materials and Methods

Results and discussion

Conclusion