Methods Used for Detection and Recovery of Escherichia coli O157:H7 Associated with a Food-Borne Disease Outbreak†

Abstract

The current Food Safety and Inspection Service method for detection and recovery of Escherichia coli O157:H7, (including modified EC broth with novobiocin (mEC+n) and a direct blot ELISA). was used to analyze beef and environmental samples during an investigation of a food-borne disease outbreak attributed to consumption of undercooked hamburger patties. Double-modified trypticase soy broth (dmTSB) and a commercially available dipstick immunoassay were also used to improve detection/recovery of E. coli O157:H7. A total of 1,115 beef and environmental samples was screened with the direct blot ELISA and the dipstick immunoassay; 178 presumptive-positive samples (by either or both of the screening methods) were subjected to recovery/isolation procedures. Toxigenic E. coli O157:H7 was recovered from 45 samples: 40 hamburger-patty samples produced on the epidemiologically identified date, 3 hamburger-patty samples produced on another date, and 2 beef briskets. The organism was not recovered from environmental samples. Limited quantitative analyses indicated that contaminated hamburger patties contained fewer than 4.3 CFU of E. coli O157:H7 per g. Atypical, toxigenic ornithine decarboxylase–negative E. coli O157:H7 and nontoxigenic sorbitol-positive E. coli O157:H29 were also recovered. Both enrichment broths gave strong positive reactions with the two immunoassay screening methods, but E. coli O157:H7 was recovered more often from mEC+n broth than from dmTSB. Both screening methods gave positive results for 44 of the 45 beef samples found to contain E. coli O157:H7. False-positive results were frequently observed with both screening methods.