Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species

Detalhes bibliográficos
Autor(a) principal: Graça, Grazielle Cardoso da
Data de Publicação: 2012
Outros Autores: Volpini, Angela Cristina, Romero, Gustavo Adolfo Sierra, Oliveira Neto, Manoel Paes de, Hueb, Marcia, Porrozzi, Renato, Boité, Mariana Côrtes, Cupolillo, Elisa
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/7478
Resumo: FIOCRUZ, FAPERJ, CNPq (MCT/CNPq/CT-Saúde/MS/SCTIE/DECIT 34/2008 - Doenças Negligenciadas - 576104/2008-6), Edital Pró-Equipamentos CAPES (27/2010), European Union (LeishEpinetSA, INCO-CT2005-015407) GC da G and ACV contributed equally to this work.
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spelling Graça, Grazielle Cardoso daVolpini, Angela CristinaRomero, Gustavo Adolfo SierraOliveira Neto, Manoel Paes deHueb, MarciaPorrozzi, RenatoBoité, Mariana CôrtesCupolillo, Elisa2014-04-01T13:05:21Z2014-04-01T13:05:21Z2012GRAÇA, Grazielle Cardoso da et al. Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Memórias do Instituto Oswaldo Cruz. Rio de Janeiro, v. 107, n. 5, p. 664-674, 2012.0074-0276https://www.arca.fiocruz.br/handle/icict/747810.1590/S0074-02762012000500014FIOCRUZ, FAPERJ, CNPq (MCT/CNPq/CT-Saúde/MS/SCTIE/DECIT 34/2008 - Doenças Negligenciadas - 576104/2008-6), Edital Pró-Equipamentos CAPES (27/2010), European Union (LeishEpinetSA, INCO-CT2005-015407) GC da G and ACV contributed equally to this work.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Grupo de Genômica e Biologia Computacional. Belo Horizonte, MG, Brasil.Universidade de Brasília. Núcleo de Medicina Tropical. Laboratório de Leishmanioses Brasília, DF, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Rio de Janeiro, RJ, Brasil.Universidade Federal de Mato Grosso. Faculdade de Medicina. Cuiabá, MT, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Coleção de Leishmania. Rio de Janeiro, RJ, Brasil.In this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorfism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.engFundação Oswaldo CruzDevelopment and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite speciesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleLeishmaniasisMolecular diagnosisSpecies identificationPolymerase chain reactionRFLPshsp70info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINALDevelopment and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species.pdfDevelopment and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species.pdfapplication/pdf2104701https://www.arca.fiocruz.br/bitstream/icict/7478/1/Development%20and%20validation%20of%20PCR-based%20assays%20for%20diagnosis%20of%20American%20cutaneous%20leishmaniasis%20and%20identification%20of%20the%20parasite%20species.pdf028cfa7410d4bb33f8abb8cf042c26cdMD51LICENSElicense.txtlicense.txttext/plain; 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dc.title.pt_BR.fl_str_mv Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
title Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
spellingShingle Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
Graça, Grazielle Cardoso da
Leishmaniasis
Molecular diagnosis
Species identification
Polymerase chain reaction
RFLPs
hsp70
title_short Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
title_full Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
title_fullStr Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
title_full_unstemmed Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
title_sort Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
author Graça, Grazielle Cardoso da
author_facet Graça, Grazielle Cardoso da
Volpini, Angela Cristina
Romero, Gustavo Adolfo Sierra
Oliveira Neto, Manoel Paes de
Hueb, Marcia
Porrozzi, Renato
Boité, Mariana Côrtes
Cupolillo, Elisa
author_role author
author2 Volpini, Angela Cristina
Romero, Gustavo Adolfo Sierra
Oliveira Neto, Manoel Paes de
Hueb, Marcia
Porrozzi, Renato
Boité, Mariana Côrtes
Cupolillo, Elisa
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Graça, Grazielle Cardoso da
Volpini, Angela Cristina
Romero, Gustavo Adolfo Sierra
Oliveira Neto, Manoel Paes de
Hueb, Marcia
Porrozzi, Renato
Boité, Mariana Côrtes
Cupolillo, Elisa
dc.subject.en.pt_BR.fl_str_mv Leishmaniasis
Molecular diagnosis
Species identification
Polymerase chain reaction
RFLPs
hsp70
topic Leishmaniasis
Molecular diagnosis
Species identification
Polymerase chain reaction
RFLPs
hsp70
description FIOCRUZ, FAPERJ, CNPq (MCT/CNPq/CT-Saúde/MS/SCTIE/DECIT 34/2008 - Doenças Negligenciadas - 576104/2008-6), Edital Pró-Equipamentos CAPES (27/2010), European Union (LeishEpinetSA, INCO-CT2005-015407) GC da G and ACV contributed equally to this work.
publishDate 2012
dc.date.issued.fl_str_mv 2012
dc.date.accessioned.fl_str_mv 2014-04-01T13:05:21Z
dc.date.available.fl_str_mv 2014-04-01T13:05:21Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv GRAÇA, Grazielle Cardoso da et al. Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Memórias do Instituto Oswaldo Cruz. Rio de Janeiro, v. 107, n. 5, p. 664-674, 2012.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/7478
dc.identifier.issn.none.fl_str_mv 0074-0276
dc.identifier.doi.none.fl_str_mv 10.1590/S0074-02762012000500014
identifier_str_mv GRAÇA, Grazielle Cardoso da et al. Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Memórias do Instituto Oswaldo Cruz. Rio de Janeiro, v. 107, n. 5, p. 664-674, 2012.
0074-0276
10.1590/S0074-02762012000500014
url https://www.arca.fiocruz.br/handle/icict/7478
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Fundação Oswaldo Cruz
publisher.none.fl_str_mv Fundação Oswaldo Cruz
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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