Department of Medical Laboratory Techniques, Nasiriyah Technical Institute, Southern Technical University, Nasiriyah, 64001, Iraq.
Department of Nursing Techniques, Nasiriyah Technical Institute, Southern Technical University, Nasiriyah, 64001, Iraq.
Acta Parasitol. 2024 Mar;69(1):831-838. doi: 10.1007/s11686-024-00817-y. Epub 2024 Mar 4.
Cutaneous leishmaniasis (CL) is still considered to be an uncontrolled endemic disease that spreads in many countries. The current study aimed to determine intra-species relationships of L. major using ITS2 sequencing.
The study was conducted from the beginning of March to the end of November 2022. All medical information regarding CL was collected from patients of Thi-Qar province who attended the Dermatology Department of Al-Hussein Teaching Hospital in Nasiriyah city. Seventy-three samples were selected for the molecular identification after confirming microscopy with Giemsa stain. In this study, the primers were designed using NCBI GenBank sequence database and Primer 3 plus primer design online software.
The results recorded 21 (28.77%) positive samples of L. major using the internal transcribed spacer 2 region (ITS2) in ribosomal RNA gene. The local L. major IQN.1-IQN.10 were submitted to NCBI GenBank database with accession numbers OM069357.1-OM069366.1, respectively. The phylogenetic analysis revealed that local isolates of L. major showed a close relationship with NCBI-BLAST L. major Iran isolate (KU680848.1).
ITS2-PCR is suitable for identifying Leishmania spp. and determining genetic diversity. A phylogenetic data analysis may provide an idea on the genetic homogeneity of local isolates and knowing the genetic origin of the dermal lesion. However, the local isolates showed genetic proximity to the KU680848.1 isolate. This signifies the possibility of infection prevalence from Iranian areas. In general, genetic variation of L. major isolates may give several clinical manifestations of the cutaneous lesion. Therefore, determination of the heterogeneity is important for detecting the infection origin, epidemiology, therapy, and control strategies.
皮肤利什曼病（CL）仍被认为是一种不受控制的地方性疾病，在许多国家传播。本研究旨在通过 ITS2 测序确定 L. major 的种内关系。
该研究于 2022 年 3 月初至 11 月底进行。从纳希里耶市侯赛因教学医院皮肤科就诊的提卡省 CL 患者中收集了所有有关 CL 的医学信息。在使用吉姆萨染色确认显微镜检查后，选择了 73 个样本进行分子鉴定。在这项研究中，使用 NCBI GenBank 序列数据库和 Primer 3 plus 引物设计在线软件设计了引物。
使用核糖体 RNA 基因内转录间隔区 2 区（ITS2）记录了 21 个（28.77%）L. major 阳性样本。本地 L. major IQN.1-IQN.10 已提交给 NCBI GenBank 数据库，登录号分别为 OM069357.1-OM069366.1。系统发育分析显示，本地 L. major 分离株与 NCBI-BLAST L. major Iran 分离株（KU680848.1）密切相关。
ITS2-PCR 适用于鉴定利什曼原虫属和确定遗传多样性。系统发育数据分析可以提供有关本地分离株遗传同质性的想法，并了解皮肤病变的遗传起源。然而，本地分离株与 KU680848.1 分离株的遗传关系较近。这表明存在从伊朗地区感染流行的可能性。一般来说，L. major 分离株的遗传变异可能导致皮肤病变的几种临床表现。因此，确定异质性对于检测感染来源、流行病学、治疗和控制策略非常重要。
