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...
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Format: | Article |
Language: | English |
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Sociedade Brasileira de Medicina Tropical (SBMT)
2015-10-01
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Series: | Revista da Sociedade Brasileira de Medicina Tropical |
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Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822015000500555&lng=en&tlng=en |
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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 |
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