Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections

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theresa.j.ochoa@uth.tmc.edu
Detalles Bibliográficos
Autores: Barletta, Francesca, Ochoa, Theresa J., Mercado, Erik H., Ruiz, Joaquim, Ecker, Lucie, Lopez, Giovanni, Mispireta, Monica, Gil, Ana I., Lanata, Claudio F., Cleary, Thomas G.
Formato: artículo
Fecha de Publicación:2015
Institución:Universidad Peruana de Ciencias Aplicadas
Repositorio:UPC-Institucional
Lenguaje:inglés
OAI Identifier:oai:repositorioacademico.upc.edu.pe:10757/556075
Enlace del recurso:http://hdl.handle.net/10757/556075
Nivel de acceso:acceso abierto
Materia:Asymptomatic Diseases
Bacterial Load
Cohort Studies
Diarrhea
Enteropathogenic Escherichia coli/
Escherichia coli Infections
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dc.title.es_PE.fl_str_mv Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
title Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
spellingShingle Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
Barletta, Francesca
Asymptomatic Diseases
Bacterial Load
Cohort Studies
Diarrhea
Enteropathogenic Escherichia coli/
Escherichia coli Infections
title_short Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
title_full Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
title_fullStr Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
title_full_unstemmed Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
title_sort Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
author Barletta, Francesca
author_facet Barletta, Francesca
Ochoa, Theresa J.
Mercado, Erik H.
Ruiz, Joaquim
Ecker, Lucie
Lopez, Giovanni
Mispireta, Monica
Gil, Ana I.
Lanata, Claudio F.
Cleary, Thomas G.
author_role author
author2 Ochoa, Theresa J.
Mercado, Erik H.
Ruiz, Joaquim
Ecker, Lucie
Lopez, Giovanni
Mispireta, Monica
Gil, Ana I.
Lanata, Claudio F.
Cleary, Thomas G.
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Barletta, Francesca
Ochoa, Theresa J.
Mercado, Erik H.
Ruiz, Joaquim
Ecker, Lucie
Lopez, Giovanni
Mispireta, Monica
Gil, Ana I.
Lanata, Claudio F.
Cleary, Thomas G.
dc.subject.es_PE.fl_str_mv Asymptomatic Diseases
Bacterial Load
Cohort Studies
Diarrhea
Enteropathogenic Escherichia coli/
Escherichia coli Infections
topic Asymptomatic Diseases
Bacterial Load
Cohort Studies
Diarrhea
Enteropathogenic Escherichia coli/
Escherichia coli Infections
description theresa.j.ochoa@uth.tmc.edu
publishDate 2015
dc.date.accessioned.es_PE.fl_str_mv 2015-05-30T17:26:05Z
dc.date.available.es_PE.fl_str_mv 2015-05-30T17:26:05Z
dc.date.issued.fl_str_mv 2015-05-30
dc.type.es_PE.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.issn.es_PE.fl_str_mv 1058-4838
dc.identifier.pmid.es_PE.fl_str_mv 22028433
dc.identifier.doi.es_PE.fl_str_mv 10.1086/648069.
dc.identifier.uri.es_PE.fl_str_mv http://hdl.handle.net/10757/556075
dc.identifier.eissn.es_PE.fl_str_mv 1537-6591
dc.identifier.journal.es_PE.fl_str_mv Clinical Infectious Diseases (Clin Infect Dis)
dc.identifier.pmcid.es_PE.fl_str_mv PMC3214587
identifier_str_mv 1058-4838
22028433
10.1086/648069.
1537-6591
Clinical Infectious Diseases (Clin Infect Dis)
PMC3214587
url http://hdl.handle.net/10757/556075
dc.language.iso.es_PE.fl_str_mv eng
language eng
dc.relation.url.es_PE.fl_str_mv http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3214587/
dc.rights.es_PE.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.es_PE.fl_str_mv application/pdf
dc.publisher.es_PE.fl_str_mv Oxford University Press
dc.source.es_PE.fl_str_mv Universidad Peruana de Ciencias Aplicadas (UPC)
Repositorio Académico - UPC
dc.source.none.fl_str_mv reponame:UPC-Institucional
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instacron:UPC
instname_str Universidad Peruana de Ciencias Aplicadas
instacron_str UPC
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spelling Barletta, FrancescaOchoa, Theresa J.Mercado, Erik H.Ruiz, JoaquimEcker, LucieLopez, GiovanniMispireta, MonicaGil, Ana I.Lanata, Claudio F.Cleary, Thomas G.2015-05-30T17:26:05Z2015-05-30T17:26:05Z2015-05-301058-48382202843310.1086/648069.http://hdl.handle.net/10757/5560751537-6591Clinical Infectious Diseases (Clin Infect Dis)PMC3214587theresa.j.ochoa@uth.tmc.eduArticleBACKGROUND: Enteropathogenic Escherichia coli (EPEC) strains are pediatric pathogens commonly isolated from both healthy and sick children with diarrhea in areas of endemicity. The aim of this study was to compare the bacterial load of EPEC isolated from stool samples from children with and without diarrhea to determine whether bacterial load might be a useful tool for further study of this phenomenon. METHODS: EPEC was detected by polymerase chain reaction (PCR) of colonies isolated on MacConkey plates from 53 diarrheal and 90 healthy children aged <2 years. DNA was isolated from stool samples by cetyltrimethylammonium bromide extraction. To standardize quantification by quantitative real-time PCR (qRT-PCR), the correlation between fluorescence threshold cycle and copy number of the intimin gene of EPEC E2348/69 was determined. RESULTS: The detection limit of qRT-PCR was 5 bacteria/mg stool. The geometric mean load in diarrhea was 299 bacteria/mg (95% confidence interval [CI], 77-1164 bacteria/mg), compared with 29 bacteria/mg (95% CI, 10-87 bacteria/mg) in control subjects (P = .016). Bacterial load was significantly higher in children with diarrhea than in control subjects among children <12 months of age (178 vs 5 bacteria/mg; P = .006) and among children with EPEC as the sole pathogen (463 vs 24 bacteria/mg; P = .006). CONCLUSIONS: EPEC load measured by qRT-PCR is higher in diarrheal than in healthy children. qRT-PCR may be useful to study the relationship between disease and colonization in settings of endemicity.Financial support. This work was supported by a Public Health Service award (grants 1K01TW007405 to T. J. O. and R01-HD051716 to T. G. C.) from the National Institutes of Health; by Agencia Espan˜ola de Cooperacio´n Internacional para el Desarrollo, Spain (D/019499/08 and D/024648/09 to J. R. and T. J. O.); and by Dr Lanata’s Institutional Research Funds. Potential conflicts of interest. All authors: No reported conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed.Revisión por paresapplication/pdfengOxford University Presshttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3214587/info:eu-repo/semantics/openAccessUniversidad Peruana de Ciencias Aplicadas (UPC)Repositorio Académico - UPCreponame:UPC-Institucionalinstname:Universidad Peruana de Ciencias Aplicadasinstacron:UPCAsymptomatic Diseases2517d314-3f5e-42be-a4e6-adbf31308569600Bacterial Load436ea583-ec62-4deb-9f57-899ec91f6bd6600Cohort Studiesfeee59d0-7546-4245-9e73-37685b0d4c5d600Diarrhea743dd4de-8a3c-4d82-9a8a-5644c228c88a600Enteropathogenic Escherichia coli/0f63c2c1-cba3-4f99-923e-0d013694e707600Escherichia coli Infections9338da0d-c0a6-4d2c-821c-a594fc2bf7ba600Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infectionsinfo:eu-repo/semantics/article2018-06-16T05:33:27ZBACKGROUND: Enteropathogenic Escherichia coli (EPEC) strains are pediatric pathogens commonly isolated from both healthy and sick children with diarrhea in areas of endemicity. The aim of this study was to compare the bacterial load of EPEC isolated from stool samples from children with and without diarrhea to determine whether bacterial load might be a useful tool for further study of this phenomenon. METHODS: EPEC was detected by polymerase chain reaction (PCR) of colonies isolated on MacConkey plates from 53 diarrheal and 90 healthy children aged <2 years. DNA was isolated from stool samples by cetyltrimethylammonium bromide extraction. To standardize quantification by quantitative real-time PCR (qRT-PCR), the correlation between fluorescence threshold cycle and copy number of the intimin gene of EPEC E2348/69 was determined. RESULTS: The detection limit of qRT-PCR was 5 bacteria/mg stool. The geometric mean load in diarrhea was 299 bacteria/mg (95% confidence interval [CI], 77-1164 bacteria/mg), compared with 29 bacteria/mg (95% CI, 10-87 bacteria/mg) in control subjects (P = .016). Bacterial load was significantly higher in children with diarrhea than in control subjects among children <12 months of age (178 vs 5 bacteria/mg; P = .006) and among children with EPEC as the sole pathogen (463 vs 24 bacteria/mg; P = .006). CONCLUSIONS: EPEC load measured by qRT-PCR is higher in diarrheal than in healthy children. qRT-PCR may be useful to study the relationship between disease and colonization in settings of endemicity.ORIGINALClin Infect Dis.-2011-Barletta-1223-9.pdfClin Infect Dis.-2011-Barletta-1223-9.pdfapplication/pdf348251https://repositorioacademico.upc.edu.pe/bitstream/10757/556075/1/Clin%20Infect%20Dis.-2011-Barletta-1223-9.pdfa4027f9c9d384d8330c3a8ac3279cae5MD51trueCC-LICENSElicense_urllicense_urltext/plain; charset=utf-849https://repositorioacademico.upc.edu.pe/bitstream/10757/556075/2/license_url4afdbb8c545fd630ea7db775da747b2fMD52falselicense_textlicense_texttext/html; charset=utf-818695https://repositorioacademico.upc.edu.pe/bitstream/10757/556075/3/license_text706fa86f01594bbd4ec888ac35076615MD53falselicense_rdflicense_rdfapplication/rdf+xml; charset=utf-823148https://repositorioacademico.upc.edu.pe/bitstream/10757/556075/4/license_rdf9da0b6dfac957114c6a7714714b86306MD54falseLICENSElicense.txtlicense.txttext/plain; charset=utf-81659https://repositorioacademico.upc.edu.pe/bitstream/10757/556075/5/license.txt1ed8f33c5404431ad7aabc05080746c5MD55falseTEXTClin Infect Dis.-2011-Barletta-1223-9.pdf.txtClin Infect Dis.-2011-Barletta-1223-9.pdf.txtExtracted Texttext/plain33344https://repositorioacademico.upc.edu.pe/bitstream/10757/556075/6/Clin%20Infect%20Dis.-2011-Barletta-1223-9.pdf.txt2dcce009156ebf530e01861761315b58MD56falseTHUMBNAILClin Infect Dis.-2011-Barletta-1223-9.pdf.jpgClin Infect Dis.-2011-Barletta-1223-9.pdf.jpgGenerated Thumbnailimage/jpeg98828https://repositorioacademico.upc.edu.pe/bitstream/10757/556075/7/Clin%20Infect%20Dis.-2011-Barletta-1223-9.pdf.jpg7a4999853e07241e931b6a4fc655baadMD57false10757/556075oai:repositorioacademico.upc.edu.pe:10757/5560752019-08-30 07:57:39.823Repositorio académico upcupc@openrepository.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Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
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