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首页> 外文期刊>Parasitology >Heat-shock protein 70 PCR-RFLP: a universal simple tool for Leishmania species discrimination in the New and Old World.
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Heat-shock protein 70 PCR-RFLP: a universal simple tool for Leishmania species discrimination in the New and Old World.

机译：热激蛋白70 PCR-RFLP：在新旧世界中用于鉴别利什曼原虫种类的通用简单工具。

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INTRODUCTION: Species typing in leishmaniasis gains importance in diagnostics, epidemiology, and clinical studies. A restriction fragment length polymorphism (RFLP) assay of PCR amplicons from a partial heat-shock protein 70 gene (hsp70) had been described for the New World, allowing identification of some species. METHODS: Based on an initial in silico analysis of 51 hsp70 sequences, most of which we recently determined in the frame of a phylogenetic study, species-specific restriction sites were identified. These were tested by PCR-RFLP on 139 strains from 14 species, thereby documenting both inter- and intra-species variability. RESULTS: Our assay could identify Leishmania infantum, L. donovani, L. tropica, L. aethiopica, L. major, L. lainsoni, L. naiffi, L. braziliensis, L. peruviana, L. guyanensis, and L. panamensis by applying 2 subsequent digests. L. mexicana, L. amazonensis, and L. garnhami did not generate species-specific restriction fragment patterns. CONCLUSION: Currently no assay is available for global Leishmania species discrimination. We present a universal PCR-RFLP method allowing identification of most medically relevant Old and New World Leishmania species on the basis of a single PCR, obviating the need to perform separate PCRs. The technique is simple to perform and can be implemented in all settings where PCR is available.
机译：简介：输入利什曼病的物种在诊断，流行病学和临床研究中具有重要意义。对于新大陆，已经描述了来自部分热休克蛋白70基因（hsp70）的PCR扩增子的限制性片段长度多态性（RFLP）分析，从而可以鉴定一些物种。方法：基于对51个hsp70序列的初步计算机分析，我们最近在系统发育研究框架中确定了其中的大多数，鉴定了物种特异性限制性位点。通过PCR-RFLP对来自14个物种的139个菌株进行了测试，从而记录了物种间和物种内的变异性。结果：我们的测定方法可以通过以下方法鉴定婴儿利什曼原虫，多诺尼乳杆菌，热带香豆乳，aethiopica乳杆菌，马来酸乳杆菌，lainsoni乳杆菌，naiffi乳杆菌，巴西乳杆菌，peruviana乳杆菌，guyanensis乳杆菌和panamensis乳杆菌。应用2个后续摘要。 L. mexicana，L. amazonensis和L. garnhami不会生成特定于物种的限制片段模式。结论：目前尚无针对全球利什曼原虫种歧视的检测方法。我们提出了一种通用的PCR-RFLP方法，可在单个PCR的基础上鉴定大多数医学上相关的旧世界和新世界利什曼原虫物种，从而无需执行单独的PCR。该技术易于执行，可在所有可用PCR的环境中实施。

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