title: Detection of E. coli O157:H7 in raw ground beef by Pathatrix™ immunomagnetic-separation, real-time PCR and cultural methods.
abstract: Detection of Escherichia coli O157:H7 by conventional cultural methods can be difficult in food matrices with a high background flora such as raw ground beef. Raw ground beef samples, artificially contaminated separately with five strains of E. coli O157:H7 at low (~0.2 cfu/g) and high (~2 cfu/g) levels, were enriched by two enrichment protocols; buffered peptone water (BPW) at 37 °C for 5h and 24h and modified buffered peptone water with pyruvate (mBPWp) for 5h at 37 °C followed by adding selective agents and incubating at 42 °C to 24h. Detection of added E. coli O157:H7 by real-time PCR (RTiPCR) and recovery on isolation agars was performed before and after PATHATRIX™ immunomagnetic separation (IMS). RTiPCR detection and cultural recovery of inoculated E. coli O157:H7 after 5h enrichment were poor at 0.21-0.24 cfu/g. The addition of IMS after 5h enrichment did not improve RTiPCR detection but markedly improved recovery by culturing. By extending enrichment to 24h, RTiPCR detection improved to 76% for either enrichment protocol without IMS. When 24h enrichment was followed by IMS, RTiPCR detection was also further improved. Cultural recovery after 24h enrichment was 56% and 84% without IMS and 100% and 92% after IMS for BPW and mBPWp respectively. Extended enrichment to 24h followed by IMS was found to be sensitive and reliable for detection and cultural recovery from raw ground beef using either enrichment method.
authors.affiliation: New Mexico State University, Food Safety Laboratory, Las Cruces, NM 88003, USA. wfedio@nmsu.edu
authors.forename: Willis M
authors.initials: WM
authors.lastname: Fedio
authors.forename: Karen C
authors.initials: KC
authors.lastname: Jinneman
authors.forename: Ken J
authors.initials: KJ
authors.lastname: Yoshitomi
authors.forename: Ruben
authors.initials: R
authors.lastname: Zapata
authors.forename: Chitra N
authors.initials: CN
authors.lastname: Wendakoon
authors.forename: Paul
authors.initials: P
authors.lastname: Browning
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Weagant
mesh_terms: D018829:Adhesins, Bacterial; D000362:Agar; D000818:Animals; D002417:Cattle; D015169:Colony Count, Microbial; D019453:Escherichia coli O157 / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology / Q000379:methods*; D018189:Immunomagnetic Separation / Q000379:methods; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D018829:Adhesins, Bacterial; D000362:Agar