Montalvo Ana M, Fraga Jorge, Tirado Dídier, Blandón Gustavo, Alba Annia, Van der Auwera Gert, Vélez Iván Darío, Muskus Carlos
Department of Parasitology, Institute of Tropical Medicine Pedro Kourí, Havana, Cuba.
PECET (Program of Study and Control of Tropical Diseases) University of Antioquia, Medellín, Colombia.
Parasitol Res. 2017 Jul;116(7):1843-1848. doi: 10.1007/s00436-017-5454-6. Epub 2017 Jun 1.
Leishmaniasis is highly prevalent in New World countries, where several methods are available for detection and identification of Leishmania spp. Two hsp70-based PCR protocols (PCR-N and PCR-F) and their corresponding restriction fragment length polymorphisms (RFLP) were applied for detection and identification of Leishmania spp. in clinical samples recruited in Colombia, Guatemala, and Honduras. A total of 93 cases were studied. The samples were classified into positive or suspected of leishmaniasis according to parasitological criteria. Molecular amplification of two different hsp70 gene fragments and further RFLP analysis for identification of Leishmania species was done. The detection in parasitologically positive samples was higher using PCR-N than PCR-F. In the total of samples studied, the main species identified were Leishmania panamensis, Leishmania braziliensis, and Leishmania infantum (chagasi). Although RFLP-N was more efficient for the identification, RFLP-F is necessary for discrimination between L. panamensis and Leishmania guyanesis, of great importance in Colombia. Unexpectedly, one sample from this country revealed an RFLP pattern corresponding to Leishmania naiffi. Both molecular variants are applicable for the study of clinical samples originated in Colombia, Honduras, and Guatemala. Choosing the better tool for each setting depends on the species circulating. More studies are needed to confirm the presence of L. naiffi in Colombian territory.
利什曼病在新大陆国家高度流行，在这些国家有多种检测和鉴定利什曼原虫属的方法。两种基于热休克蛋白70（hsp70）的聚合酶链反应（PCR）方案（PCR - N和PCR - F）及其相应的限制性片段长度多态性（RFLP）被应用于检测和鉴定从哥伦比亚、危地马拉和洪都拉斯收集的临床样本中的利什曼原虫属。共研究了93例病例。根据寄生虫学标准，将样本分为利什曼病阳性或疑似病例。对两个不同的hsp70基因片段进行分子扩增，并进一步进行RFLP分析以鉴定利什曼原虫种类。使用PCR - N在寄生虫学阳性样本中的检测率高于PCR - F。在所研究的样本中，鉴定出的主要种类为巴拿马利什曼原虫、巴西利什曼原虫和婴儿利什曼原虫（恰加斯亚种）。虽然RFLP - N在鉴定方面更有效，但RFLP - F对于区分巴拿马利什曼原虫和圭亚那利什曼原虫是必要的，这在哥伦比亚非常重要。出乎意料的是，来自该国的一个样本显示出与奈菲利什曼原虫相对应的RFLP模式。这两种分子变体都适用于研究源自哥伦比亚、洪都拉斯和危地马拉的临床样本。为每种情况选择更好的工具取决于流行的物种。需要更多研究来证实奈菲利什曼原虫在哥伦比亚境内的存在。
