title: Rapid categorization of pathogenic Escherichia coli by multiplex PCR.
abstract: A one-shot multiplex polymerase chain reaction (PCR) was developed for detecting 12 virulence genes of diarrheagenic Escherichia coli. In order to differentiate between the five categories of diarrheagenic E. coli, we selected the target genes: stx1, stx2, and eaeA for enterohemorrhagic E. coli(EHEC); eaeA, bfpA, and EAF for enteropathogenic E. coli(EPEC); invE for enteroinvasive E. coli(EIEC); elt, estp, and esth for enterotoxigenic E. coli(ETEC); CVD432 and aggR for enteroaggregative E. coli(EAggEC); and astA distributed over the categories of diarrheagenic E. coli. In our multiplex PCR system, all 12 targeted genes (stx1, stx2, eaeA, invE, elt, estp, astA, esth, bfpA, aggR, EAF, and CVD432) were amplified in a single PCR reaction in one tube and detected by electrophoresis. Using our multiplex PCR, the 208 clinically isolated strains of diarrheagenic E. coli in our laboratory were successfully categorized and easily analyzed for the presence of virulence plasmids.
authors.affiliation: Department of Bacteriology, Toyama Institute of Health, Japan. keiko.kimata@pref.toyama.lg.jp
authors.forename: Keiko
authors.initials: K
authors.lastname: Kimata
authors.forename: Tomoko
authors.initials: T
authors.lastname: Shima
authors.forename: Miwako
authors.initials: M
authors.lastname: Shimizu
authors.forename: Daisuke
authors.initials: D
authors.lastname: Tanaka
authors.forename: Junko
authors.initials: J
authors.lastname: Isobe
authors.forename: Yotaku
authors.initials: Y
authors.lastname: Gyobu
authors.forename: Masanori
authors.initials: M
authors.lastname: Watahiki
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Nagai
mesh_terms: D004926:Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000382:microbiology*; D005243:Feces / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014774:Virulence