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

dc.creatorBarletta, Francesca
dc.creatorOchoa, Theresa J.
dc.creatorMercado, Erik H.
dc.creatorRuiz, Joaquim
dc.creatorEcker, Lucie
dc.creatorLopez, Giovanni
dc.creatorMispireta, Monica
dc.creatorGil, Ana I.
dc.creatorLanata, Claudio F.
dc.creatorCleary, Thomas G.
dc.date2015-05-30
dc.date.accessioned2017-03-02T19:54:07Z
dc.date.available2017-03-02T19:54:07Z
dc.identifier1058-4838
dc.identifier22028433
dc.identifier10.1086/648069.
dc.identifierhttp://hdl.handle.net/10757/556075
dc.identifierhttp://repositorioacademico.upc.edu.pe/upc/handle/10757/556075
dc.identifier1537-6591
dc.identifierClinical Infectious Diseases (Clin Infect Dis)
dc.identifierPMC3214587
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/169101
dc.descriptiontheresa.j.ochoa@uth.tmc.edu
dc.descriptionArticle
dc.descriptionBACKGROUND: 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.
dc.languageeng
dc.publisherOxford University Press
dc.relationhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3214587/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectAsymptomatic Diseases
dc.subjectBacterial Load
dc.subjectCohort Studies
dc.subjectDiarrhea
dc.subjectEnteropathogenic Escherichia coli/
dc.subjectEscherichia coli Infections
dc.titleQuantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
dc.typeArtículos de revistas


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