Koarashi Yu, Cáceres Abraham G, Saca Florencia Margarita Zúniga, Flores Elsa Elvira Palacios, Trujillo Adela Celis, Alvares José Luis Abanto, Yoshimatsu Kumiko, Arikawa Jiro, Katakura Ken, Hashiguchi Yoshihisa, Kato Hirotomo
Laboratory of Parasitology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan; Department of Microbiology, Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
Sección de Entomología, Instituto de Medicina Tropical "Daniel A. Carrión" y Departamento Académico de Microbiología Médica, Facultad de Medicina Humana, Universidad Nacional Mayor de San Marcos, Lima, Peru; Laboratorio de Entomología, Instituto Nacional de Salud, Lima, Peru.
Acta Trop. 2016 Jun;158:83-87. doi: 10.1016/j.actatropica.2016.02.024. Epub 2016 Mar 2.
A PCR-Restriction Fragment Length Polymorphism (RFLP) targeting the mannose phosphate isomerase gene was established to differentiate Leishmania species distributed near the Department of Huanuco, Peru. The technique was applied to 267 DNA samples extracted from Giemsa-stained smears of cutaneous lesions taken from patients suspected for cutaneous leishmaniasis in the area, and the present status of causative Leishmania species was identified. Of 114 PCR-amplified samples, 22, 19, 24 and 49 samples were identified to be infected by Leishmania (Viannia) braziliensis, L. (V.) peruviana, L. (V.) guyanensis, and a hybrid of L. (V.) braziliensis/L. (V.) peruviana, respectively, and the validity of PCR-RFLP was confirmed by sequence analysis. Since PCR-RFLP is simple and rapid, the technique will be a useful tool for the epidemiological study of leishmaniasis.
建立了一种针对磷酸甘露糖异构酶基因的聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法，用于区分秘鲁瓦努科省附近分布的利什曼原虫种类。该技术应用于从该地区疑似皮肤利什曼病患者的皮肤病变吉姆萨染色涂片提取的267份DNA样本，并确定了致病利什曼原虫种类的现状。在114份PCR扩增样本中，分别有22份、19份、24份和49份样本被鉴定为感染了巴西利什曼原虫（维阿尼亚种）、秘鲁利什曼原虫（维阿尼亚种）、圭亚那利什曼原虫（维阿尼亚种）以及巴西利什曼原虫（维阿尼亚种）/秘鲁利什曼原虫（维阿尼亚种）的杂交种，并且通过序列分析证实了PCR-RFLP的有效性。由于PCR-RFLP简单快速，该技术将成为利什曼病流行病学研究的有用工具。
