Aghaei Afshar A, Rassi Y, Sharifi I, Vatandoost H, Mollaie Hr, Oshaghi M A, Abai Mr, Rafizadeh S
Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran.
Department of Medical Entomology & Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Asian Pac J Trop Med. 2014 Feb;7(2):93-6. doi: 10.1016/S1995-7645(14)60002-X.
OBJECTIVE: To identify the Leishmania species in infected sand flies by Real-time PCR coupled with HRM analysis. METHODS: Real-time PCR coupled with HRM analysis targeting the first internal transcribed spacer (ITS1) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish Leishmania species in sand flies specimens. RESULTS: Three out of 115 females of Phlebotomus sergenti (P. sergenti) (2.6%) were positive to Leishmania tropica (L. tropica). CONCLUSIONS: This is the first report on P. sergenti as the main and proven vector of anthroponitic cutaneous leishmaniasis in Dehbakri County using Real-time PCR coupled with HRM analysis. This method is rapid, sensitive and specific for diagnosing of parasites in infected Sand flies and ideal for large scale genotyping projects.
目的：通过实时荧光定量聚合酶链反应（Real-time PCR）结合高分辨率熔解曲线分析（HRM）来鉴定感染白蛉体内的利什曼原虫种类。 方法：以核糖体DNA的第一内部转录间隔区（ITS1）为遗传标记，采用实时荧光定量聚合酶链反应结合高分辨率熔解曲线分析，对白蛉标本中的利什曼原虫种类进行鉴定和区分。 结果：115只中华白蛉雌蛉中有3只（2.6%）对热带利什曼原虫呈阳性反应。 结论：这是首次报道在德哈卜克里县使用实时荧光定量聚合酶链反应结合高分辨率熔解曲线分析，证实中华白蛉是皮肤利什曼病的主要传播媒介。该方法对于诊断感染白蛉体内的寄生虫快速、灵敏且特异，是大规模基因分型项目的理想选择。
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