Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin

ABSTRACTINTRODUCTION: 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 Leishmaniadeoxyr...

Full description

Bibliographic Details
Main Authors: 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
Format: Article
Language:English
Published: Sociedade Brasileira de Medicina Tropical (SBMT) 2015-10-01
Series:Revista da Sociedade Brasileira de Medicina Tropical
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822015000500555&lng=en&tlng=en
_version_ 1818233357313507328
author 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
author_facet 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
author_sort Suzane Ribeiro Prestes
collection DOAJ
description ABSTRACTINTRODUCTION: 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.
first_indexed 2024-12-12T11:20:54Z
format Article
id doaj.art-83e1bb8846db4dbabb1ba32915af096c
institution Directory Open Access Journal
issn 1678-9849
language English
last_indexed 2024-12-12T11:20:54Z
publishDate 2015-10-01
publisher Sociedade Brasileira de Medicina Tropical (SBMT)
record_format Article
series Revista da Sociedade Brasileira de Medicina Tropical
spelling doaj.art-83e1bb8846db4dbabb1ba32915af096c2022-12-22T00:26:02ZengSociedade Brasileira de Medicina Tropical (SBMT)Revista da Sociedade Brasileira de Medicina Tropical1678-98492015-10-0148555555910.1590/0037-8682-0132-2015S0037-86822015000500555Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffinSuzane Ribeiro PrestesJorge Augusto de Oliveira GuerraGustavo Adolfo Sierra RomeroLaylah Kelre Costa MagalhaesRosa Amelia Gonçalves SantanaMarcel Gonçalves MacielAna CustódioMaria das Graças Vale BarbosaHenrique SilveiraABSTRACTINTRODUCTION: 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822015000500555&lng=en&tlng=enPCRLeishmania braziliensisLeishmania guyanensisMucosal leishmaniasisMolecular diagnosis
spellingShingle 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
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
PCR
Leishmania braziliensis
Leishmania guyanensis
Mucosal leishmaniasis
Molecular diagnosis
title Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin
title_full Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin
title_fullStr Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin
title_full_unstemmed Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin
title_short Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin
title_sort polymerase chain reaction based method for the identification of leishmania viannia braziliensis and leishmania viannia guyanensis in mucosal tissues conserved in paraffin
topic PCR
Leishmania braziliensis
Leishmania guyanensis
Mucosal leishmaniasis
Molecular diagnosis
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822015000500555&lng=en&tlng=en
work_keys_str_mv AT suzaneribeiroprestes polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT jorgeaugustodeoliveiraguerra polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT gustavoadolfosierraromero polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT laylahkelrecostamagalhaes polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT rosaameliagoncalvessantana polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT marcelgoncalvesmaciel polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT anacustodio polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT mariadasgracasvalebarbosa polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin
AT henriquesilveira polymerasechainreactionbasedmethodfortheidentificationofleishmaniavianniabraziliensisandleishmaniavianniaguyanensisinmucosaltissuesconservedinparaffin