Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis

Detalhes bibliográficos
Autor(a) principal: Waggoner, Jesse J
Data de Publicação: 2014
Outros Autores: Balassiano, Ilana, Abeynayake, Janaki, Sahoo, Malaya K, Mohamed-Hadley, Alisha, Liu, Yuanyuan, Vita-Brasil, Juliana Magalhães, Pinsky, Benjamin A
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/10866
Resumo: Stanford University School of Medicine. Division of Infectious Diseases and Geographic Medicine. Department of Medicine. Stanford, CA, USA.
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spelling Waggoner, Jesse JBalassiano, IlanaAbeynayake, JanakiSahoo, Malaya KMohamed-Hadley, AlishaLiu, YuanyuanVita-Brasil, Juliana MagalhãesPinsky, Benjamin A2015-06-17T12:05:20Z2015-06-17T12:05:20Z2014WAGGONER, Jesse J. et al. Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis. Plos One, v.9, n.11, 8p, 2014.https://www.arca.fiocruz.br/handle/icict/1086610.1371/journal.pone.0112356engPlos OneSensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleStanford University School of Medicine. Division of Infectious Diseases and Geographic Medicine. Department of Medicine. Stanford, CA, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. WHO/PAHO Centro Colaborador para Leptospirose. Coleção de Leptospira. Centro de Referência Nacional para Leptospirose. Laboratório de Zoonoses Bacterianas. Rio de Janeiro, RJ, Brasil.Stanford University School of Medicine. Department of Pathology. Stanford, CA, USA.Stanford University School of Medicine. Department of Pathology. Stanford, CA, USA.Stanford University School of Medicine. Department of Pathology. Stanford, CA, USA.Stanford University School of Medicine. Department of Pathology. Stanford, CA, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. WHO/PAHO Centro Colaborador para Leptospirose. Coleção de Leptospira. Centro de Referência Nacional para Leptospirose. Laboratório de Zoonoses Bacterianas. Rio de Janeiro, RJ, Brasil.Stanford University School of Medicine. Division of Infectious Diseases and Geographic Medicine. Department of Medicine. Department of Pathology. Stanford, CA, USA.Background: Bacteria of the genus Leptospira, the causative agents of leptospirosis, are categorized into pathogenic and non-pathogenic species. However, the benefit of using a clinical diagnostic that is specific for pathogenic species remains unclear. In this study, we present the development of a real-time PCR (rtPCR) for the detection of pathogenic Leptospira (the pathogenic rtPCR), and we perform a comparison of the pathogenic rtPCR with a published assay that detects all Leptospira species [the undifferentiated febrile illness (UFI) assay] and a reference 16S Leptospira rtPCR, which was originally designed to detect pathogenic species. Methodology/Principal Findings: For the pathogenic rtPCR, a new hydrolysis probe was designed for use with primers from the UFI assay, which targets the 16S gene. The pathogenic rtPCR detected Leptospira DNA in 37/37 cultured isolates from 5 pathogenic and one intermediate species. Two strains of the non-pathogenic L. biflexa produced no signal. Clinical samples from 65 patients with suspected leptospirosis were then tested using the pathogenic rtPCR and a reference Leptospira 16S rtPCR. All 65 samples had tested positive for Leptospira using the UFI assay; 62 (95.4%) samples tested positive using the pathogenic rtPCR (p = 0.24). Only 24 (36.9%) samples tested positive in the reference 16S rtPCR (p,0.0001 for comparison with the pathogenic rtPCR and UFI assays). Amplicon sequencing confirmed the detection of pathogenic Leptospira species in 49/50 cases, including 3 cases that were only detected using the UFI assay. Conclusions/Significance: The pathogenic rtPCR displayed similar sensitivity to the UFI assay when testing clinical specimens with no difference in specificity. Both assays proved significantly more sensitive than a real-time molecular test used for comparison. Future studies are needed to investigate the clinical and epidemiologic significance of more sensitive Leptospira detection using these tests.LeptospirosisPCRLeptospiraLeptospiroseReação em Cadeia da Polimeraseinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txttext/plain1914https://www.arca.fiocruz.br/bitstream/icict/10866/1/license.txt7d48279ffeed55da8dfe2f8e81f3b81fMD51ORIGINALilana_balassiano2etal_IOC_2014.pdfapplication/pdf564052https://www.arca.fiocruz.br/bitstream/icict/10866/2/ilana_balassiano2etal_IOC_2014.pdf80570b9162fb99d11eb720869f4a7548MD52TEXTilana_balassiano2etal_IOC_2014.pdf.txtilana_balassiano2etal_IOC_2014.pdf.txtExtracted texttext/plain39948https://www.arca.fiocruz.br/bitstream/icict/10866/3/ilana_balassiano2etal_IOC_2014.pdf.txta37664b48f5ade49f72c5da863a99599MD53icict/108662022-06-24 13:12:38.093oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352022-06-24T16:12:38Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
title Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
spellingShingle Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
Waggoner, Jesse J
Leptospirosis
PCR
Leptospira
Leptospirose
Reação em Cadeia da Polimerase
title_short Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
title_full Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
title_fullStr Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
title_full_unstemmed Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
title_sort Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis
author Waggoner, Jesse J
author_facet Waggoner, Jesse J
Balassiano, Ilana
Abeynayake, Janaki
Sahoo, Malaya K
Mohamed-Hadley, Alisha
Liu, Yuanyuan
Vita-Brasil, Juliana Magalhães
Pinsky, Benjamin A
author_role author
author2 Balassiano, Ilana
Abeynayake, Janaki
Sahoo, Malaya K
Mohamed-Hadley, Alisha
Liu, Yuanyuan
Vita-Brasil, Juliana Magalhães
Pinsky, Benjamin A
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Waggoner, Jesse J
Balassiano, Ilana
Abeynayake, Janaki
Sahoo, Malaya K
Mohamed-Hadley, Alisha
Liu, Yuanyuan
Vita-Brasil, Juliana Magalhães
Pinsky, Benjamin A
dc.subject.en.pt_BR.fl_str_mv Leptospirosis
PCR
Leptospira
topic Leptospirosis
PCR
Leptospira
Leptospirose
Reação em Cadeia da Polimerase
dc.subject.decs.pt_BR.fl_str_mv Leptospirose
Reação em Cadeia da Polimerase
description Stanford University School of Medicine. Division of Infectious Diseases and Geographic Medicine. Department of Medicine. Stanford, CA, USA.
publishDate 2014
dc.date.issued.fl_str_mv 2014
dc.date.accessioned.fl_str_mv 2015-06-17T12:05:20Z
dc.date.available.fl_str_mv 2015-06-17T12:05:20Z
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 WAGGONER, Jesse J. et al. Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis. Plos One, v.9, n.11, 8p, 2014.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/10866
dc.identifier.doi.pt_BR.fl_str_mv 10.1371/journal.pone.0112356
identifier_str_mv WAGGONER, Jesse J. et al. Sensitive Real-Time PCR Detection of Pathogenic Leptospira spp. and a Comparison of Nucleic Acid Amplification Methods for the Diagnosis of Leptospirosis. Plos One, v.9, n.11, 8p, 2014.
10.1371/journal.pone.0112356
url https://www.arca.fiocruz.br/handle/icict/10866
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.none.fl_str_mv Plos One
publisher.none.fl_str_mv Plos One
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)
collection Repositório Institucional da FIOCRUZ (ARCA)
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