The PCR-based detection and identification of the parasites causing human cutaneous leishmaniasis in the Iranian city of Ahvaz.

M Ghasemian, S Maraghi, A R Samarbafzadeh, A Jelowdar, M Kalantari
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引用次数: 51

Abstract

In Iran, Leishmania major or L. tropica cause almost all of the human cutaneous leishmaniasis (CL). Unfortunately, the detection methods frequently used for CL (the microscopical examination of direct smears or the culture of biopsies) are not very sensitive and the Leishmania species causing each case of CL in Iran is usually only tentatively identified from extrinsic factors, such as the case's clinical manifestations and region of residence. Recently, however, a nested PCR that targets the parasites' kinetoplast DNA has been used in the city of Ahvaz (the capital of the province of Khouzestan, in south-western Iran) to confirm the microscopical diagnosis of CL and to identify the causative parasites, to species level. Smears from the lesions on 100 suspected cases of CL were fixed, stained with Wright's eosin-methylene blue, and checked for amastigotes under a light microscope. Scrapings from the same smears were then tested for leishmanial DNA, using a nested PCR that allows the DNA from L. tropica to be identified and distinguished from that of L. major. The 100 smears investigated were all found amastigote-positive by microscopy and PCR-positive for either L. major DNA (97 smears) or L. tropica DNA (three smears). The predominant species causing CL in Ahvaz is therefore L. major.

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伊朗阿瓦士市致人皮肤利什曼病寄生虫的pcr检测和鉴定
在伊朗,大利什曼原虫或热带利什曼原虫引起几乎所有的人类皮肤利什曼病(CL)。不幸的是,经常用于CL的检测方法(直接涂片的显微镜检查或活检培养)不是很敏感,并且在伊朗引起每个CL病例的利什曼原虫物种通常只能从外部因素(如病例的临床表现和居住地区)初步确定。然而,最近在Ahvaz市(伊朗西南部Khouzestan省的首府)使用了一种针对寄生虫着丝体DNA的巢式PCR,以确认CL的显微镜诊断并在物种水平上鉴定致病寄生虫。对100例疑似CL病例的病变涂片进行固定,用怀特氏伊红-亚甲基蓝染色,并在光镜下检查有无无刚体。然后,使用巢式PCR对来自相同涂片的刮屑进行利什曼DNA测试,该方法可以识别和区分来自热带乳杆菌的DNA和主要乳杆菌的DNA。100份涂片镜检均发现无梭菌阳性,pcr阳性主要乳杆菌DNA(97份涂片)或热带乳杆菌DNA(3份涂片)。因此,在阿瓦士造成CL的主要物种是L. major。
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Annals of tropical medicine and parasitology
Annals of tropical medicine and parasitology 医学-公共卫生、环境卫生与职业卫生
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