LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools

Detalhes bibliográficos
Autor(a) principal: Santos, Helena Lúcia Carneiro
Data de Publicação: 2013
Outros Autores: Bandyopadhyay, Kalali, Bandea, Rebecca, Peralta, Regina Helena Saramago, Peralta, José Mauro, Silva, Alexandre Januário da
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/11828
Resumo: Universidade Federal do Rio de Janeiro. Instituto de Microbiologia. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. laboratório de Avaliação e Promoção da Saúde Ambiental. Rio de Janeiro, RJ, Brasil.
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spelling Santos, Helena Lúcia CarneiroBandyopadhyay, KalaliBandea, RebeccaPeralta, Regina Helena SaramagoPeralta, José MauroSilva, Alexandre Januário da2015-09-28T13:02:35Z2015-09-28T13:02:35Z2013SANTOS, Helena Lúcia Carneiro; et al. LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools. Parasites & Vectors, v.6, n.60, 9p, 2013.1756-3305https://www.arca.fiocruz.br/handle/icict/1182810.1186/1756-3305-6-69engBioMed CentralLUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stoolsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversidade Federal do Rio de Janeiro. Instituto de Microbiologia. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. laboratório de Avaliação e Promoção da Saúde Ambiental. Rio de Janeiro, RJ, Brasil.Center for Global Health. Division of Parasitic Diseases and Malaria. Centers for Disease Control and Prevention. Atlanta, GA, USA.Center for Global Health. Division of Parasitic Diseases and Malaria. Centers for Disease Control and Prevention. Atlanta, GA, USA.Universidade Federal Fluminense. Departamento de Patologia. Niterói, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Microbiologia. Rio de Janeiro, RJ, Brasil.Center for Global Health. Division of Parasitic Diseases and Malaria. Centers for Disease Control and Prevention. Atlanta, GA, USA.Background: Six species of the genus Entamoeba, i.e., E. histolytica, E. dispar, E. moshkovskii, E. polecki, E. coli, and E. hartmanii can be found in human stools. Among these, only E. histolytica is considered to be pathogenic, causing intestinal and extra-intestinal disease, but it is morphologically identical to E. dispar and E. moshkovskii. In general, E. polecki, E. coli, and E. hartmanii can be differentiated morphologically from E. histolytica, but some of their diagnostic morphologic features may overlap creating issues for the differential diagnosis. Moreover, the previous inability to differentiate among Entamoeba species has limited epidemiologic information on E histolytica. The objective of this study was to develop a rapid, high-throughput screening method using Luminex technique for the simultaneous detection and differentiation of Entamoeba species. Methods: PCR amplification was performed with biotinylated Entamoeba sp 18S rRNA gene primers, designed to amplify a fragment ranging from 382 to 429 bp of the Entamoeba spp studied. Regions of this fragment that could differentiate among E. histolytica, E. moshkovskii, E. dispar, E. hartmanii and E. coli were selected to design hybridization probes to link to Luminex beads. The assay was standardized with cloned DNA samples of each species and evaluated with 24 DNA extracts from samples obtained from individuals diagnosed with these amebas in their stools. Results: Using this approach we were able to correctly identify E. histoltyica, E. dispar, E hartmanni, E. coli and E. moshkovskii in all specimens studied. From twenty four samples tested by microscopy, PCR/DNA Sequencing and real-time PCR, 100% agreed with PCR-Luminex assay for identification of E. dispar, E. moshkovskii, E. hartmanni, E. histolytica, and E. coli. Conclusion: These results show that this method could be used in the diagnostic detection of Entamoeba spp in fecal samples. This diagnostic test was useful to clearly distinguish E histolytica from other species and also to strengthen epidemiologic data on Entamoeba spp.Entamoeba18SrRNAPCRMolecular DiagnosisMultiplex AssayEntamoebaReação em Cadeia da PolimerasePatologia Molecularinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txttext/plain1914https://www.arca.fiocruz.br/bitstream/icict/11828/1/license.txt7d48279ffeed55da8dfe2f8e81f3b81fMD51ORIGINALhelena_santos_etal_IOC_2013.pdfapplication/pdf373011https://www.arca.fiocruz.br/bitstream/icict/11828/2/helena_santos_etal_IOC_2013.pdfc537be69eb18cae08af818dd30e1222aMD52TEXThelena_santos_etal_IOC_2013.pdf.txthelena_santos_etal_IOC_2013.pdf.txtExtracted texttext/plain46879https://www.arca.fiocruz.br/bitstream/icict/11828/3/helena_santos_etal_IOC_2013.pdf.txtf465260f7bbe178eec4ba616a5c78959MD53icict/118282022-06-24 13:10:51.077oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352022-06-24T16:10:51Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
title LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
spellingShingle LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
Santos, Helena Lúcia Carneiro
Entamoeba
18SrRNA
PCR
Molecular Diagnosis
Multiplex Assay
Entamoeba
Reação em Cadeia da Polimerase
Patologia Molecular
title_short LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
title_full LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
title_fullStr LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
title_full_unstemmed LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
title_sort LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools
author Santos, Helena Lúcia Carneiro
author_facet Santos, Helena Lúcia Carneiro
Bandyopadhyay, Kalali
Bandea, Rebecca
Peralta, Regina Helena Saramago
Peralta, José Mauro
Silva, Alexandre Januário da
author_role author
author2 Bandyopadhyay, Kalali
Bandea, Rebecca
Peralta, Regina Helena Saramago
Peralta, José Mauro
Silva, Alexandre Januário da
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Santos, Helena Lúcia Carneiro
Bandyopadhyay, Kalali
Bandea, Rebecca
Peralta, Regina Helena Saramago
Peralta, José Mauro
Silva, Alexandre Januário da
dc.subject.en.pt_BR.fl_str_mv Entamoeba
18SrRNA
PCR
Molecular Diagnosis
Multiplex Assay
topic Entamoeba
18SrRNA
PCR
Molecular Diagnosis
Multiplex Assay
Entamoeba
Reação em Cadeia da Polimerase
Patologia Molecular
dc.subject.decs.pt_BR.fl_str_mv Entamoeba
Reação em Cadeia da Polimerase
Patologia Molecular
description Universidade Federal do Rio de Janeiro. Instituto de Microbiologia. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. laboratório de Avaliação e Promoção da Saúde Ambiental. Rio de Janeiro, RJ, Brasil.
publishDate 2013
dc.date.issued.fl_str_mv 2013
dc.date.accessioned.fl_str_mv 2015-09-28T13:02:35Z
dc.date.available.fl_str_mv 2015-09-28T13:02:35Z
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 SANTOS, Helena Lúcia Carneiro; et al. LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools. Parasites & Vectors, v.6, n.60, 9p, 2013.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/11828
dc.identifier.issn.pt_BR.fl_str_mv 1756-3305
dc.identifier.doi.pt_BR.fl_str_mv 10.1186/1756-3305-6-69
identifier_str_mv SANTOS, Helena Lúcia Carneiro; et al. LUMINEXW: a new technology for the simultaneous identification of five Entamoeba spp. commonly found in human stools. Parasites & Vectors, v.6, n.60, 9p, 2013.
1756-3305
10.1186/1756-3305-6-69
url https://www.arca.fiocruz.br/handle/icict/11828
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 BioMed Central
publisher.none.fl_str_mv BioMed Central
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)
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