Evaluation of a multiplex PCR method to detect enteroaggregative Escherichia coli

Biocell. 2006 Aug;30(2):301-8.

Abstract

Enteroaggregative Escherichia coli (EAEC) has been implicated in sporadic diarrhea in children and adults and has been identified as the cause of several outbreaks worldwide. The HEp-2 test remains the gold standard for identification of this pathotype. A 60-65 MDa plasmid encodes the aggregative adherence fimbriae (AAF/I and AAF/II), a transcriptional activator (aggR gene), the enteroaggregative heat-stable enterotoxin EAST1 (astA gene) and a cytotoxin (Pet). The standard assay for EAEC is performed only in research laboratories, because it is expensive, labor intensive and time-consuming. The Polymerase Chain Reaction (PCR) offers the possibility of rapid diagnosis. In the current study, a multiplex PCR assay which checks aggR and astA genes was designed. Eigthy-eight E. coli strains, isolated from children with acute diarrhea in Mendoza, Argentina, were characterized by the reference method (HEp-2 assay), and by aggR-astA PCR. A strong correlation between the presence of the specific marker aggR and the reference test was found. The astA gene had a similar distribution between aggregative and localized strains, indicating that this gene could not be considered as a marker of EAEC. We conclude that aggR may be used to identify EAEC, using the PCR method as a screening test.

MeSH terms

  • Bacterial Adhesion / physiology
  • DNA, Bacterial / analysis
  • DNA, Bacterial / genetics
  • Electrophoresis, Agar Gel
  • Escherichia coli / cytology
  • Escherichia coli / genetics*
  • Escherichia coli / isolation & purification*
  • Escherichia coli Proteins / genetics
  • Humans
  • Infant
  • Polymerase Chain Reaction / methods*
  • Serotyping
  • Trans-Activators / genetics

Substances

  • AggR protein, E coli
  • DNA, Bacterial
  • Escherichia coli Proteins
  • Trans-Activators