title: Detection of Shiga toxin-producing Escherichia coli O26, O45, O103, O111, O113, O121, O145, and O157 serogroups by multiplex polymerase chain reaction of the wzx gene of the O-antigen gene cluster.
abstract: O-antigens on the surface of Escherichia coli are important virulence factors that are targets of both the innate and adaptive immune system and play a major role in pathogenicity. O-antigens that are responsible for antigenic specificity of the strain determine the O-serogroup. E. coli O26, O45, O103, O111, O113, O121, O145, and O157 have been the most commonly identified O-serogroups associated with Shiga toxin-producing E. coli (STEC) implicated in outbreaks of human illness all over the world. A multiplex polymerase chain reaction assay was developed to simultaneously detect the eight STEC O-serogroups targeting the wzx (O-antigen-flippase) genes of all O-antigen gene clusters. The sensitivity of the multiplex polymerase chain reaction was found to be 10 colony forming units for each O-group when enriched in broth and 100 colony forming units when enriched in artificially inoculated apple juice diluted with tryptic soy broth for 16 h at 37°C. The method can be used for detecting STEC O-groups simultaneously and may be exploited for improving the safety of food products.
authors.affiliation: E. coli Reference Center, Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, Wiley Lane, University Park, PA 16802, USA. rcd3@psu.edu
authors.forename: Chitrita
authors.initials: C
authors.lastname: DebRoy
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Roberts
authors.forename: Angela M
authors.initials: AM
authors.lastname: Valadez
authors.forename: Edward G
authors.initials: EG
authors.lastname: Dudley
authors.forename: Catherine N
authors.initials: CN
authors.lastname: Cutter
mesh_terms: D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics*; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction; D019081:O Antigens / Q000235:genetics; D012680:Sensitivity and Specificity; D054323:Shiga-Toxigenic Escherichia coli / Q000145:classification* / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D019081:O Antigens; C543926:Wzx protein, E coli