title: Multiplex real-time PCR (MRT-PCR) for diarrheagenic.
abstract: Diarrheagenic Escherichia coli strains are important causes of diarrhea in children from the developing world and are now being recognized as emerging enteropathogens in the developed world. Current methods of detection are too expensive and labor-intensive for routine detection of these organisms to be practical. We developed a real-time fluorescence-based multiplex PCR for the detection of all six of the currently recognized classes of diarrheagenic E. coli. The primers were designed to specifically amplify eight different virulence genes in the same reaction: aggR for enteroaggregative E. coli (EAEC), stIa/stIb and lt for enterotoxigenic E. coli (ETEC), eaeA for enteropathogenic E. coli (EPEC), stx1 and stx2 for Shiga toxin-producing E. coli (STEC), ipaH for enteroinvasive E. coli (EIEC), and daaD for diffusely adherent E. coli (DAEC).
authors.affiliation: Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru.
authors.forename: Francesca
authors.initials: F
authors.lastname: Barletta
authors.forename: Theresa J
authors.initials: TJ
authors.lastname: Ochoa
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Cleary
mesh_terms: D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D003967:Diarrhea / Q000175:diagnosis* / Q000382:microbiology*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000175:diagnosis*; D005243:Feces / Q000382:microbiology; D005798:Genes, Bacterial; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D044366:Transition Temperature
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial