Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections
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Authors
Barletta, FrancescaOchoa, Theresa J.
Mercado, Erik H.
Ruiz, Joaquim
Ecker, Lucie
Lopez, Giovanni
Mispireta, Monica
Gil, Ana I.
Lanata, Claudio F.
Cleary, Thomas G.
Issue Date
2015-05-30Keywords
Asymptomatic DiseasesBacterial Load
Cohort Studies
Diarrhea
Enteropathogenic Escherichia coli/
Escherichia coli Infections
Metadata
Show full item recordPublisher
Oxford University PressJournal
Clinical Infectious Diseases (Clin Infect Dis)DOI
10.1086/648069.PubMed ID
22028433PubMed Central ID
PMC3214587Additional Links
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3214587/Abstract
BACKGROUND: 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.Type
info:eu-repo/semantics/articleRights
info:eu-repo/semantics/openAccessLanguage
engISSN
1058-4838EISSN
1537-6591ae974a485f413a2113503eed53cd6c53
10.1086/648069.
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