Laboratório de Pesquisa em Leishmaniose, Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil 4365, Manguinhos, 21045-900 Rio de Janeiro, RJ, Brazil.
Infect Genet Evol. 2010 Jan;10(1):77-83. doi: 10.1016/j.meegid.2009.11.001. Epub 2009 Nov 11.
Since the first report of the genus Leishmania, many species have been described. In Brazil, human leishmaniasis has been associated with eight Leishmania species, of which seven are responsible for cutaneous leishmaniasis (CL). In some endemic areas, CL is associated with only one species; however, in other areas, such as the Amazon, the etiology of CL can be assigned to many species. The multitude of highly similar Leishmania species in Brazil makes it difficult to develop an appropriate method of typing them. Most Leishmania species were first described based on epidemiological and biological characteristics, and these were later corroborated by Multilocus Enzyme Electrophoresis (MLEE), the gold standard technique for identifying Leishmania species. In an attempt to overcome the limitations of MLEE, many PCR-based methods have been developed and used for parasite identification. In the present study, we analyzed the sequence of the hsp70 gene in Leishmania species associated with human leishmaniasis in Brazil. This analysis led to the identification of restriction enzymes that could be used for PCR-RFLP-based identification. The results obtained were in complete agreement with those obtained by MLEE, suggesting that PCR-RFLP analysis of hsp70 could soon replace MLEE for routine Leishmania typing.
自利什曼原虫属的第一个报告以来，已经描述了许多物种。在巴西，人类利什曼病与八种利什曼原虫有关，其中七种可导致皮肤利什曼病（CL）。在一些流行地区，CL 仅与一种物种有关；然而，在其他地区，如亚马逊地区，CL 的病因可归因于多种物种。巴西存在许多高度相似的利什曼原虫物种，这使得对其进行适当的分型方法变得困难。大多数利什曼原虫物种最初是根据流行病学和生物学特征进行描述的，后来通过多基因座酶电泳（MLEE）进行了证实，这是鉴定利什曼原虫物种的金标准技术。为了克服 MLEE 的局限性，已经开发并使用了许多基于 PCR 的方法来进行寄生虫鉴定。在本研究中，我们分析了与巴西人类利什曼病相关的利什曼原虫种的 hsp70 基因序列。该分析导致鉴定出可用于基于 PCR-RFLP 的鉴定的限制酶。所得结果与 MLEE 完全一致，表明 hsp70 的 PCR-RFLP 分析很快可能取代 MLEE 用于常规利什曼原虫分型。
