Prestes Suzane Ribeiro, Guerra Jorge Augusto de Oliveira, Romero Gustavo Adolfo Sierra, Magalhaes Laylah Kelre Costa, Santana Rosa Amelia Gonçalves, Maciel Marcel Gonçalves, Custódio Ana, Barbosa Maria das Graças Vale, Silveira Henrique
Universidade do Estado do Amazonas, Manaus, Amazonas, Brazil.
Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Amazonas, Brazil.
Rev Soc Bras Med Trop. 2015 Sep-Oct;48(5):555-9. doi: 10.1590/0037-8682-0132-2015.
In the Americas, mucosal leishmaniasis is primarily associated with infection by Leishmania (Viannia) braziliensis. However, Leishmania (Viannia) guyanensis is another important cause of this disease in the Brazilian Amazon. In this study, we aimed at detecting Leishmaniadeoxyribonucleic acid (DNA) within paraffin-embedded fragments of mucosal tissues, and characterizing the infecting parasite species.
We evaluated samples collected from 114 patients treated at a reference center in the Brazilian Amazon by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses.
Direct examination of biopsy imprints detected parasites in 10 of the 114 samples, while evaluation of hematoxylin and eosin-stained slides detected amastigotes in an additional 17 samples. Meanwhile, 31/114 samples (27.2%) were positive for Leishmania spp. kinetoplast deoxyribonucleic acid (kDNA) by PCR analysis. Of these, 17 (54.8%) yielded amplification of the mini-exon PCR target, thereby allowing for PCR-RFLP-based identification. Six of the samples were identified as L. (V.) braziliensis, while the remaining 11 were identified as L. (V.) guyanensis.
The results of this study demonstrate the feasibility of applying molecular techniques for the diagnosis of human parasites within paraffin-embedded tissues. Moreover, our findings confirm that L. (V.) guyanensisis a relevant causative agent of mucosal leishmaniasis in the Brazilian Amazon.
在美洲，黏膜利什曼病主要与巴西利什曼原虫（维阿尼亚种）感染有关。然而，圭亚那利什曼原虫（维阿尼亚种）是巴西亚马逊地区该疾病的另一个重要病因。在本研究中，我们旨在检测黏膜组织石蜡包埋片段中的利什曼原虫脱氧核糖核酸（DNA），并鉴定感染的寄生虫种类。
我们通过聚合酶链反应（PCR）和限制性片段长度多态性（RFLP）分析，评估了从巴西亚马逊地区一家参考中心接受治疗的114例患者采集的样本。
对活检印片的直接检查在114个样本中的10个中检测到寄生虫，而苏木精和伊红染色玻片评估在另外17个样本中检测到无鞭毛体。同时，通过PCR分析，114个样本中有31个（27.2%）利什曼原虫属动基体脱氧核糖核酸（kDNA）呈阳性。其中，17个（54.8%）产生了微小外显子PCR靶点的扩增，从而能够基于PCR-RFLP进行鉴定。6个样本被鉴定为巴西利什曼原虫（维阿尼亚种），其余11个被鉴定为圭亚那利什曼原虫（维阿尼亚种）。
本研究结果证明了应用分子技术诊断石蜡包埋组织中人体寄生虫的可行性。此外，我们的研究结果证实，圭亚那利什曼原虫（维阿尼亚种）是巴西亚马逊地区黏膜利什曼病的相关病原体。
