Quantitative Real-time Polymerase Chain Reaction for Enteropathogenic Escherichia coli: A Tool for Investigation of Asymptomatic Versus Symptomatic Infections
Year: 2011
Type: article
Study design: Observational Study
Abstract: 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.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.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).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.
Source: Greater South Information System
DOI: 10.60692/8hjz3-50879
Authors: Francesca Barletta, Theresa J. Ochoa, Erik H. Mercado, Joaquı́m Ruiz, Lucie Ecker, Gladys Galvis López, Mónica Mispireta, Ana I. Gil, Claudio F. Lanata, Thomas G. Cleary
Institutions: Universidad Peruana Cayetano Heredia, The University of Texas Health Science Center at Houston, Centro de Investigación Biomédica en Red de Epidemiología y Salud Pública, Universitat de Barcelona, Instituto de Investigación Nutricional, Idaho State University, Peruvian University of Applied Sciences
Language: English
Subfield: Endocrinology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2231430668