title: Development and assessment of a rapid method to detect Escherichia coli O26, O111 and O157 in retail minced beef.
abstract: A molecular-based detection method was developed to detect Escherichia coli O26, O111 and O157 in minced (ground) beef samples. This method consists of an initial overnight enrichment in modified tryptone soya broth (mTSB) and novobiocin prior to DNA extraction and subsequent serogrouping using a triplex PCR. This method has a low limit of detection and results are available within 24 hours of receipt of samples. Once optimized, this rapid method was utilized to determine the prevalence of these E. coli serogroups in six hundred minced beef samples all of which were previously examined by immunomagnetic separation (IMS) and selective plating for E. coli O26 and O111. Using IMS, two E. coli O26 isolates were detected. No E. coli O111 were recovered. The multiplex PCR technique described here did not detect E. coli O111 nor O157 in any of the samples, however six minced beef samples were positive for E. coli O26 using our method, only two of these were previously detected by IMS and culture. Application of molecular methods are useful to support culture-based approaches thereby further contributing to risk reduction along the food chain.
authors.affiliation: Public Health Laboratory, Health Service Executive-South Western Area, Cherry Orchard Hospital, Ballyfermot, Dublin, Ireland.sfanning@ucd.ie
authors.forename: Mary
authors.initials: M
authors.lastname: Murphy
authors.forename: Anne
authors.initials: A
authors.lastname: Carroll
authors.forename: Ciara
authors.initials: C
authors.lastname: Walsh
authors.forename: Paul
authors.initials: P
authors.lastname: Whyte
authors.forename: Micheál
authors.initials: M
authors.lastname: O'Mahony
authors.forename: Wayne
authors.initials: W
authors.lastname: Anderson
authors.forename: Eleanor
authors.initials: E
authors.lastname: McNamara
authors.forename: Séamus
authors.initials: S
authors.lastname: Fanning
mesh_terms: D000818:Animals; D002417:Cattle; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D005511:Food Handling; D008460:Meat / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial