ABSTRACT
BACKGROUND: Childhood diarrheal diseases remain highly endemic in developing areas of Brazil. The importance of Escherichia coli among children with diarrhea in these areas was unknown. This study determined the prevalence of different E. coli categories in symptomatic and asymptomatic children from low socioeconomic level rural communities in southeastern Brazil. METHODS: A total of 560 stool samples were collected from 141 children with diarrhea (< 10 years) and 419 apparently healthy controls who resided in 23 communities. E. coli isolates (n = 1943) were subjected to two multiplex PCRs developed for the detection of enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), diffusely adherent E. coli (DAEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and Shiga toxin-producing E. coli (STEC). Strains were also examined for the presence of EPEC, EAEC, and DAEC by assays of adhesion to HEp-2 cells and by hybridization with specific DNA probes. RESULTS: Diarrheagenic E. coli strains were isolated from 253 (45.2%) children, and were associated with diarrhea in children aged < 5 years (p < 0.001). EAEC (20.9%), DAEC (11.6%), EPEC (9.3%) were the most frequent pathotypes, followed by ETEC (2.7%), EIEC (0.5%), and STEC (0.2%). Depending of the assay, EPEC, EAEC, and DAEC (collectively termed enteroadherent E. coli) strains were isolated in 45% to 56% of diarrhea cases, a significantly higher incidence than in controls (P < 0.05). Individually, only DAEC showed significant association with diarrhea (p < 0.05), particularly in children aged 2-5 years. CONCLUSION: This study indicates that enteroadherent E. coli is an important cause of diarrhea in children living in low socioeconomic level communities in southeastern Brazil. Our results reveal that the PCR1 assay is an excellent tool for the identification of EAEC and DAEC.
Subject(s)
Diarrhea/microbiology , Enteropathogenic Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Brazil/epidemiology , Child, Preschool , Diarrhea/diagnosis , Diarrhea/economics , Diarrhea/epidemiology , Enteropathogenic Escherichia coli/genetics , Escherichia coli Infections/diagnosis , Escherichia coli Infections/economics , Escherichia coli Infections/epidemiology , Feces/microbiology , Female , Genotype , Humans , Incidence , Infant , Male , Phenotype , PovertyABSTRACT
A real-time multiplex PCR assay was designed to amplify the virulence genes eae, pEAF, aatA, daaC, elt, est, ipaH, stx(1), and stx(2) for the detection of all diarrheagenic Escherichia coli pathotypes. This assay proved to be more sensitive and rapid than a conventional multiplex PCR for diarrheagenic E. coli isolates from children with diarrhea.
