Detection of Escherichia coli serogroups O26 and O113 by PCR amplification of the wzx and wzy genes.
PCR-based assays for detecting enterohemorrhagic Escherichia coli serogroups O26 and O113 were developed by targeting the wzx (O-antigen flippase) and the wzy (O-antigen polymerase) genes found in the O-antigen gene cluster of each organism. The PCR assays were specific for the respective serogroups, as there was no amplification of DNA from non-O26 and non-O113 E. coli serogroups or from other bacterial genera tested. Using the PCR assays, we were able to detect the organisms in seeded apple juice inoculated at concentration levels as low as < or =10 CFU/ml. The O26- and O113-specific PCR assays can potentially be used for typing E. coli O26 and O113 serogroups; these assays will offer an advantage to food and environmental microbiology laboratories in terms of identifying these non-O157 serogroups by replacing antigen-based serotyping.
Gastroenteric Disease Center, Department of Veterinary Science, The Pennsylvania State University, University Park, Pennsylvania 16802, USA. rcd3@psu.edu
Chitrita
C
DebRoy
Elisabeth
E
Roberts
James
J
Kundrat
Michael A
MA
Davis
Connie E
CE
Briggs
Pina M
PM
Fratamico
D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D005798:Genes, Bacterial; D006602:Hexosyltransferases / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D012703:Serotyping
D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; C117254:RfbX protein, Pseudomonas; D006602:Hexosyltransferases; C085652:O-antigen polymerase