Mirahmadi Hadi, Rezaee Nasrin, Mehravaran Ahmad, Heydarian Peyman, Raeghi Saber
Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran.
Department of Parasitology and Mycology, Faculty of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran.
Vet World. 2018 May;11(5):700-705. doi: 10.14202/vetworld.2018.700-705. Epub 2018 May 26.
Cutaneous leishmaniasis (CL) is one of the most important health problems that are capable of involving both tropical and subtropical areas, especially in Iran. This cross-sectional study aimed to differentiate the species that are able to cause CL in Zahedan city by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.
It was conducted on 145 suspected CL patients in Zahedan city between 2014 and 2016. The smears were initially prepared, air-dried, fixed with absolute methanol, and stained with 10% Giemsa. Then, we examined the stained samples by a light microscope under 1000× magnifications. PCR assay targeted cytochrome (cyt ) gene using LCBF1 and LCBR2 primers and the products digested by Ssp1 enzymes.
From 145 suspected CL patients, 76 (52.4%) were positive in microscopic examination. In addition, we detected gene of interest (cyt ) in 98 (67.5%). The results of PCR-RFLP indicated that 53/98 (54%) cases were and 45/98 (46%) were , and the main species in these areas was .
We concluded that the microscopic examination is not sensitive enough and is not able to distinguish between different species. Instead, molecular methods like PCR-RFLP can be appropriately used with promising results.
皮肤利什曼病（CL）是最重要的健康问题之一，在热带和亚热带地区均有发生，尤其是在伊朗。本横断面研究旨在通过聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法鉴别能够在扎黑丹市引起CL的物种。
对2014年至2016年间扎黑丹市145例疑似CL患者进行研究。涂片最初制备、风干、用无水甲醇固定，并用10%吉姆萨染色。然后，我们在1000倍放大倍数下通过光学显微镜检查染色样本。PCR检测使用LCBF1和LCBR2引物靶向细胞色素（cyt）基因，产物用Ssp1酶消化。
在145例疑似CL患者中，76例（52.4%）显微镜检查呈阳性。此外，我们在98例（67.5%）中检测到目标基因（cyt）。PCR-RFLP结果表明，98例中有53例（54%）为 ，45例（46%）为 ，这些地区的主要物种为 。
我们得出结论，显微镜检查不够灵敏，无法区分不同的 物种。相反，像PCR-RFLP这样的分子方法可以适当地使用并取得有希望的结果。
