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Title: Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin
Authors: Prestes, Suzane Ribeiro
Guerra, Jorge Augusto de Oliveira
Romero, Gustavo Adolfo Sierra
Magalhães, Laylah Kelre Costa
Santana, Rosa Amelia Gonçalves
Maciel, Marcel Gonçalves
Custódio, Ana
Barbosa, Maria das Graças Vale
Silveira, Henrique
Assunto:: Leishmaniose
Diagnóstico molecular
Issue Date: Sep-2015
Publisher: Sociedade Brasileira de Medicina Tropical - SBMT
Citation: PRESTES, Suzane Ribeiro et al. Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin. Revista da Sociedade Brasileira de Medicina Tropical, Uberaba, v. 48, n. 5, p. 555-559, set./out. 2015. Disponível em: <http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822015000500555&lng=en&nrm=iso>. Acesso em: 10 maio 2018. doi: http://dx.doi.org/10.1590/0037-8682-0132-2015.
Abstract: Introduction: 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. Methods: 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. Results: 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. Conclusions: 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.
Licença:: Revista da Sociedade Brasileira de Medicina Tropical - This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0). Fonte: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822015000500555&lng=en&nrm=iso. Acesso em: 10 maio 2018.
DOI: http://dx.doi.org/10.1590/0037-8682-0132-2015
Appears in Collections:FMD - Artigos publicados em periódicos e preprints

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