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Home / Publications / Polymerase chain reaction-based method for the identification of leishmania (viannia) Braziliensis and leishmania (viannia) guyanensis in mucosal tissues conserved in paraffin

Polymerase chain reaction-based method for the identification of leishmania (viannia) Braziliensis and leishmania (viannia) guyanensis in mucosal tissues conserved in paraffin

  • Authors: Barbosa Md, Custodio A, Guerra JA, Maciel MG, Magalhaes LK, Prestes SR, Romero GA, Santana RA, Silveira H
  • Publication Year: 2015
  • Journal: Revista da Sociedade Brasileira de Medicina Tropical
  • Link: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822015000500555&lng=en&tlng=en

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.

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About GHTM

GHTM is a R&D Unit that brings together researchers with a track record in Tropical Medicine and International & Global Health. It aims at strengthening Portugal's role as a leading partner in the development and implementation of a global health research agenda. Our evidence-based interventions contribute to the promotion of equity in health and to improve the health of populations.

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