Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.

Detalhes bibliográficos
Autor(a) principal: Backstedt, Brian T
Data de Publicação: 2015
Outros Autores: Buyuktanir, Ozlem, Lindow, Janet C, Wunder Junior, Elsio Augusto, Reis, Mitermayer Galvão dos, Usmani-Brown, Sahar, Ledizet, Michel, Ko, Albert Icksang, Pal, Utpal
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
DOI: 10.1371/journal.pone.0128913
Texto Completo: https://www.arca.fiocruz.br/handle/icict/11211
Resumo: University of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USA
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spelling Backstedt, Brian TBuyuktanir, OzlemLindow, Janet CWunder Junior, Elsio AugustoReis, Mitermayer Galvão dosUsmani-Brown, SaharLedizet, MichelKo, Albert IcksangPal, Utpal2015-07-10T14:04:19Z2015-07-10T14:04:19Z2015BACKSTEDT, B. T. et al. Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA. PLoS One, v. 10, n. 6, p. e0128913, 2015.1932-6203https://www.arca.fiocruz.br/handle/icict/1121110.1371/journal.pone.0128913engPublic Library of ScienceLeptospiroseLeptospiraSorotipagemHumanosRNA bacterianoAntígenos bacterianosEfficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversity of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USAUniversity of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USAYale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USA / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilYale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USAFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilL2 Diagnostics. New Haven, Connecticut, USAL2 Diagnostics. New Haven, Connecticut, USAYale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USA / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilUniversity of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USAPathogenic Leptospira species cause a prevalent yet neglected zoonotic disease with mild to life-threatening complications in a variety of susceptible animals and humans. Diagnosis of leptospirosis, which primarily relies on antiquated serotyping methods, is particularly challenging due to presentation of non-specific symptoms shared by other febrile illnesses, often leading to misdiagnosis. Initiation of antimicrobial therapy during early infection to prevent more serious complications of disseminated infection is often not performed because of a lack of efficient diagnostic tests. Here we report that specific regions of leptospiral 16S ribosomal RNA molecules constitute a novel and efficient diagnostic target for PCR-based detection of pathogenic Leptospira serovars. Our diagnostic test using spiked human blood was at least 100-fold more sensitive than corresponding leptospiral DNA-based quantitative PCR assays, targeting the same 16S nucleotide sequence in the RNA and DNA molecules. The sensitivity and specificity of our RNA assay against laboratory-confirmed human leptospirosis clinical samples were 64% and 100%, respectively, which was superior then an established parallel DNA detection assay. Remarkably, we discovered that 16S transcripts remain appreciably stable ex vivo, including untreated and stored human blood samples, further highlighting their use for clinical detection of L. interrogans. Together, these studies underscore a novel utility of RNA targets, specifically 16S rRNA, for development of PCR-based modalities for diagnosis of human leptospirosis, and also may serve as paradigm for detection of additional bacterial pathogens for which early diagnosis is warranted.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82354https://www.arca.fiocruz.br/bitstream/icict/11211/1/license.txt8b4c200b4e10021c5683c6ccaba07169MD51ORIGINALBackstedt BT Efficient detection....pdfBackstedt BT Efficient detection....pdfapplication/pdf1132240https://www.arca.fiocruz.br/bitstream/icict/11211/2/Backstedt%20BT%20Efficient%20detection....pdf594bed889caa4f952f8fa5e6fd1ef402MD52TEXTBackstedt BT Efficient detection....pdf.txtBackstedt BT Efficient detection....pdf.txtExtracted texttext/plain65939https://www.arca.fiocruz.br/bitstream/icict/11211/3/Backstedt%20BT%20Efficient%20detection....pdf.txt46222d04edd5faea9b918b6ce209fbcbMD53icict/112112023-03-15 14:33:06.402oai:www.arca.fiocruz.br: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Repositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352023-03-15T17:33:06Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
title Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
spellingShingle Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
Backstedt, Brian T
Leptospirose
Leptospira
Sorotipagem
Humanos
RNA bacteriano
Antígenos bacterianos
title_short Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
title_full Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
title_fullStr Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
title_full_unstemmed Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
title_sort Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA.
author Backstedt, Brian T
author_facet Backstedt, Brian T
Buyuktanir, Ozlem
Lindow, Janet C
Wunder Junior, Elsio Augusto
Reis, Mitermayer Galvão dos
Usmani-Brown, Sahar
Ledizet, Michel
Ko, Albert Icksang
Pal, Utpal
author_role author
author2 Buyuktanir, Ozlem
Lindow, Janet C
Wunder Junior, Elsio Augusto
Reis, Mitermayer Galvão dos
Usmani-Brown, Sahar
Ledizet, Michel
Ko, Albert Icksang
Pal, Utpal
author2_role author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Backstedt, Brian T
Buyuktanir, Ozlem
Lindow, Janet C
Wunder Junior, Elsio Augusto
Reis, Mitermayer Galvão dos
Usmani-Brown, Sahar
Ledizet, Michel
Ko, Albert Icksang
Pal, Utpal
dc.subject.other.pt_BR.fl_str_mv Leptospirose
Leptospira
Sorotipagem
Humanos
RNA bacteriano
Antígenos bacterianos
topic Leptospirose
Leptospira
Sorotipagem
Humanos
RNA bacteriano
Antígenos bacterianos
description University of Maryland. College Park. Department of Veterinary Medicine and Virginia-Maryland Regional College of Veterinary Medicine. Maryland, USA
publishDate 2015
dc.date.accessioned.fl_str_mv 2015-07-10T14:04:19Z
dc.date.available.fl_str_mv 2015-07-10T14:04:19Z
dc.date.issued.fl_str_mv 2015
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 BACKSTEDT, B. T. et al. Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA. PLoS One, v. 10, n. 6, p. e0128913, 2015.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/11211
dc.identifier.issn.none.fl_str_mv 1932-6203
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pone.0128913
identifier_str_mv BACKSTEDT, B. T. et al. Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA. PLoS One, v. 10, n. 6, p. e0128913, 2015.
1932-6203
10.1371/journal.pone.0128913
url https://www.arca.fiocruz.br/handle/icict/11211
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 Public Library of Science
publisher.none.fl_str_mv Public Library of Science
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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reponame_str Repositório Institucional da FIOCRUZ (ARCA)
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