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

Suzane Ribeiro Prestes, Jorge Augusto de Oliveira Guerra, Gustavo Adolfo Sierra Romero, Laylah Kelre Costa Magalhaes, Rosa Amelia Gonçalves Santana, Marcel Gonçalves Maciel, Ana Custódio, Maria das Graças Vale Barbosa, Henrique Silveira

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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 Leishmania deoxyribonucleic acid (DNA) within paraffi n-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 amplifi cation of the mini-exon PCR target, thereby allowing for PCR-RFLP-based identifi cation. Six of the samples were identifi ed as L. (V.) braziliensis, while the remaining 11 were identifi ed as L. (V.) guyanensis. Conclusions: The results of this study demonstrate the feasibility of applying molecular techniques for the diagnosis of human parasites within paraffi nembedded tissues. Moreover, our fi ndings confi rm that L. (V.) guyanensis is a relevant causative agent of mucosal leishmaniasis in the Brazilian Amazon.

Original languageEnglish
Pages (from-to)555-559
Number of pages5
JournalRevista Da Sociedade Brasileira De Medicina Tropical
Volume48
Issue number5
DOIs
Publication statusPublished - 1 Sep 2015

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Keywords

  • Leishmania Braziliensis
  • Leishmania guyanensis
  • Molecular diagnosis
  • Mucosal leishmaniasis
  • PCR

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