Koohsar Faramarz, Hajjaran Homa, Heidari Soudabeh, Darabi Enayat, Fathi Saeid, Borhani Mehdi, Mesgarian Fatemeh
Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Pour sina, Tehran 1417613151, Iran.
Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Azadi, Tehran 1419963111, Iran.
Ann Parasitol. 2020;66(3):339-346. doi: 10.17420/ap6603.272.
Cutaneous leishmaniosis (CL) is mainly caused by Leishmania major (rural-type) and Leishmania tropica (urban-type). CL is a major health problem in many regions of the world, and it is associated with health complications and economic loss. The identification and differentiation of Leishmania species are critical because the prevention and control methods, as well as management and therapeutic strategies, are different for each type of CL. The present study aimed to identify the parasite species responsible for CL in the study area using ITS1 and HSP70- based PCR-RFLP methods. A total of 147 stained slides were prepared from samples collected from CL patients, and these slides were positive for amastigotes of Leishmania species on microscopic examination. Forty-three Giemsastained slides with 2+ to 4+ grades were selected for molecular studies for the identification of the Leishmania species. DNA was extracted from the selected slides for the molecular studies. The amplification of HSP70 and ITS1 genes was performed by the PCR method. The PCR products were digested with the HaeIII restriction enzyme, and banding patterns of all samples were compared with reference strains. Overall, patterns of all the samples were found to correspond to the reference strains of L. major based on RFLP-PCR targeting HSP70 and ITS1 genes of the parasite, demonstrating the dominance of L. major as the causative agent of zoonotic cutaneous leishmaniosis (zCL) in the study area. This area is endemic for zoonotic CL, and further studies are required to determine the reservoir and natural infection of sand flies in this county.
皮肤利什曼病（CL）主要由硕大利什曼原虫（农村型）和热带利什曼原虫（城市型）引起。CL是世界许多地区的一个主要健康问题，并且与健康并发症和经济损失相关。利什曼原虫种类的鉴定和区分至关重要，因为针对每种类型的CL，其预防和控制方法以及管理和治疗策略都有所不同。本研究旨在使用基于ITS1和HSP70的PCR-RFLP方法鉴定研究区域内导致CL的寄生虫种类。从CL患者采集的样本中总共制备了147张染色玻片，这些玻片在显微镜检查下利什曼原虫无鞭毛体呈阳性。选择43张吉姆萨染色等级为2+至4+的玻片进行分子研究以鉴定利什曼原虫种类。从所选玻片中提取DNA用于分子研究。通过PCR方法扩增HSP70和ITS1基因。PCR产物用HaeIII限制酶消化，并将所有样本的条带模式与参考菌株进行比较。总体而言，基于针对寄生虫HSP70和ITS1基因的RFLP-PCR，发现所有样本的模式均与硕大利什曼原虫的参考菌株相符，表明硕大利什曼原虫作为该研究区域人畜共患皮肤利什曼病（zCL）的病原体占主导地位。该地区为人畜共患CL的流行区，需要进一步研究以确定该县白蛉的宿主和自然感染情况。
