Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species
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Publication Date: | 2012 |
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Format: | Article |
Language: | eng |
Source: | Repositório Institucional da FIOCRUZ (ARCA) |
Download full: | https://www.arca.fiocruz.br/handle/icict/7478 |
Summary: | 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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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 |
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Fundação Oswaldo Cruz (FIOCRUZ) |
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FIOCRUZ |
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FIOCRUZ |
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Repositório Institucional da FIOCRUZ (ARCA) |
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