Motalleb Gholamreza, Mirahmadi Hadi, Zare-Zadeh Ahmad, Mehravaran Ahmad
Dept. of Biology, Faculty of Sciences, University of Zabol, Zabol, Iran.
Infectious Diseases and Tropical Medicine Research Center, Resistant Tuberculosis Institute, Zahedan University of Medical Sciences, Zahedan, Iran.
Iran J Parasitol. 2017 Oct-Dec;12(4):534-543.
Despite the high prevalence and drug resistance of disease in Sistan and Baluchestan Province of Iran, the species of cutaneous leishmaniasis (CL) has not been identified. In the present study, cytochrome (Cyt ) was used in Sistan and Baluchestan to find species of in suspected patients of CL using PCR-RFLP and DNA sequencing.
This study was conducted from Oct 2015 to Oct 2016. The samples were collected from the individuals clinically suspected to CL and referred to Iran Shahr, Chabahar, Khash, Zabol, Zahedan, Mirjaveh, and Nikshahr health centers. Overall, 700 Giemsa-Stained slides from the wound of patients suspected of CL were passive collected and examined under a light microscope at ×1000. After DNA extraction, positive samples were used for Cyt detection by PCR-RFLP to determine the parasite species. One hundred positive slides were selected for molecular studies. Among positive samples, 20% were sequenced. To compare the results of sequences, molecular evolutionary genetic analysis (MEGA6) was used.
Overall, 53 samples were identified as and 47 samples (47%) . Cyt in and is converted to 400 and 480 bp and 130, 215 and 535 bp pieces respectively. In the isolated and , nucleotide changes were 3-5 (mainly in wobble site).
Infection was more related to . PCR-RFLP method has a high sensitivity for diagnosis of species.
尽管伊朗锡斯坦-俾路支斯坦省疾病的患病率高且存在耐药性，但皮肤利什曼病（CL）的种类尚未确定。在本研究中，在锡斯坦-俾路支斯坦省使用细胞色素（Cyt），通过聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）和DNA测序，在疑似CL患者中寻找利什曼原虫的种类。
本研究于2015年10月至2016年10月进行。样本取自临床疑似CL且转诊至伊朗沙赫尔、恰巴哈尔、卡什、扎博勒、扎黑丹、米尔贾韦赫和尼克沙赫尔健康中心的个体。总体而言，被动收集了700张疑似CL患者伤口的吉姆萨染色玻片，并在×1000倍光学显微镜下检查。DNA提取后，将阳性样本用于通过PCR-RFLP检测Cyt以确定寄生虫种类。选择100张阳性玻片进行分子研究。在阳性样本中，20%进行测序。为比较序列结果，使用了分子进化遗传分析（MEGA6）。
总体而言，53个样本被鉴定为杜氏利什曼原虫，47个样本（47%）为热带利什曼原虫。杜氏利什曼原虫和热带利什曼原虫中的Cyt分别转化为400和480 bp以及130、215和535 bp的片段。在分离的杜氏利什曼原虫和热带利什曼原虫中，核苷酸变化为3 - 5个（主要在摆动位点）。
感染与热带利什曼原虫的关系更为密切。PCR-RFLP方法对热带利什曼原虫种类的诊断具有高敏感性。
