DNA sequence of the Escherichia coli O103 O antigen gene cluster and detection of enterohemorrhagic E. coli O103 by PCR amplification of the wzx and wzy genes.
Escherichia coli serogroup O103 has been associated with gastrointestinal illness and hemolytic uremic syndrome. To develop PCR-based methods for detection and identification of this serogroup, the DNA sequence of the 12,033-bp region containing the O antigen gene cluster of Escherichia coli O103 was determined. Of the 12 open reading frames identified, the E. coli O103 wzx (O antigen flippase) and wzy (O antigen polymerase) genes were selected as targets for development of both conventional and real-time PCR assays specific for this serogroup. In addition, a multiplex PCR targeting the Shiga toxin (Stx) 1 (stx1), Shiga toxin 2 (stx2), wzx, and wzy genes was developed to differentiate Stx-producing E. coli O103 from non-toxigenic strains. The PCR assays can be employed to identify E. coli serogroup O103, replacing antigen-based serotyping, and to potentially detect the organism in food, fecal, or environmental samples.
Fratamico
Pina M
PM
Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor, PA 19038, USA. pfratamico@errc.ars.usda.gov
DebRoy
Chitrita
C
Strobaugh
Terence P
TP
Chen
Chin-Yi
CY
D000818:Animals; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics*; D006801:Humans; D005810:Multigene Family; D019081:O Antigens / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D022622:Shiga Toxin 1 / Q000235:genetics; D022641:Shiga Toxin 2 / Q000235:genetics; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D019081:O Antigens; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2