title: Reduction of food matrix interference by a combination of sample preparation and multi-dimensional gating techniques to facilitate rapid, high sensitivity analysis for Escherichia coli serotype O157 by flow cytometry.
abstract: Escherichia coli serotype O157 strains, which may be found in foods, often produce enterohemorrhagic toxins. The research goal was to facilitate rapid, sensitive detection in foods of E. coli serotype O157 by flow cytometry. Sample preparation methods were developed for potential use in 15 foods. Combined with multi-dimensional gating, these methods decreased time-to-results (TTR) for determination of low-level contamination. They mitigated the effects of interfering food components, concentrated cells for analysis without growth or, when necessary, used short-term incubation. The results showed qualitative analysis that was equivalent to culture plating in accuracy and superior in sensitivity and speed. Preparation time was 10-30 min per sample and detection took 3-4 min. Culture growth, if required, took an additional 4-6 h. A protocol for raw spinach analysis, using 4 h culture incubation, was 94% correct with one false negative for a low level inoculation. Its projected limit-of-detection (LOD) was 1 viable cell per 25 g of spinach, based on an average of 28 counts detected after growth and an estimated counts-to-threshold (C/T) ratio of 1.3. The results suggested potential uses for regulatory screening and food industry process control.
authors.affiliation: National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA. Jon.wilkes@fda.hhs.gov
authors.forename: Jon G
authors.initials: JG
authors.lastname: Wilkes
authors.forename: Randal K
authors.initials: RK
authors.lastname: Tucker
authors.forename: James A
authors.initials: JA
authors.lastname: Montgomery
authors.forename: Willie M
authors.initials: WM
authors.lastname: Cooper
authors.forename: John B
authors.initials: JB
authors.lastname: Sutherland
authors.forename: Dan A
authors.initials: DA
authors.lastname: Buzatu
mesh_terms: D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D019453:Escherichia coli O157 / Q000254:growth & development / Q000302:isolation & purification*; D005434:Flow Cytometry / Q000379:methods*; D005504:Food Analysis; D005506:Food Contamination / Q000032:analysis*; D005511:Food Handling / Q000379:methods; D005516:Food Microbiology / Q000379:methods*; D005638:Fruit / Q000382:microbiology; D012680:Sensitivity and Specificity; D014675:Vegetables / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial