Isolation and Identification of

E. coli O157:H7 in Foods

Conventional Methods Proposed Method
1
FDA
USDA/FSIS2 Homogenize 25 g
Homogenize sample in sample in 225 mL of
EHEC Enrichment Broth Homogenize sample in EEB1 or mECn2
(EEB - modified TSB modified EC Medium
w/o novobiocin + cefixime, w/novobiocin (mECn)2
Incubate 20 - 24 h
cefsulodin and vancomycin)1 Incubate 20 - 24 h at 35°C
at 35°C
Incubate overnight
Perform antibody-based
at 37°C
Antibody-based screening screening test or IMS
test for O157:H7/NM for O157:H7/NM as
SMAC-TC recommended1,2
(Sorbitol MacConkey Agar
w/Tellurite and Cefixime)
Positive
Negative Perform Immunomagnetic
No further testing required Separation (IMS) BBL™ CHROMagar™ O157
Incubate 18 - 24 h
at 35 - 37°C
Incubate 18 - 24 h
at 35°C
Colorless or neutral/gray Selective chromogenic
colonies w/smoky center medium for E. coli O157
on SMAC-TC Mauve-colored colonies on
Subculture to Tryptic Soy Agar Incubate 24 - 26 h BBL™ CHROMagar™ O157 are
w/0.6% Yeast Extract (TSAYE) at 35 ± 2°C presumptively identified
for confirmation as E. coli O157
Presumptive positive result Subculture to a nonselective
Subculture to a nonselective medium for confirmation
Incubate 18 - 24 h medium for confirmation
at 35°C
Incubate 18 – 24 h
at 35°C

Confirmation Tests
Rapid miniaturized test kit
or BD Products for E. coli O157 Testing in Foods
Cellobiose Fermentation
Serological Tests Indole test Cat. No. Product Pkg.
Incubate 18 – 24 h Lysine Decarboxylase 214984 BBL™ CHROMagar™ O157 20 plates
E. Coli O Antiserum O157 at 35°C 218105 Buffered Peptone Water 500 g
Motility
E. Coli H Antiserum H7 292735 Modified TSB 500 g
MR-VP Tests
Latex Kit for E. coli O157:H7 234020 Modified EC Medium 500 g
Ornithine Decarboxylase
Rapid DNA Probe test 231971 Novobiocin Antimicrobic Supplement 500 g
Simmons Citrate
Triple Sugar Iron Agar (TSI)

References
1. Feng, P. and S.D. Weagant. 2002. Diarrheagenic Escherichia coli, Chapter 4A, rev. Sept. 2002. In FDA Bacteriological analytical manual, 8th ed., Rev. A. AOAC International, Gaithersburg, MD.
2. United States Department of Agriculture, Food Safety Inspection Service. 2002. Detection, isolation and identification of Escherichia coli O157:H7 and O157:NM (Nonmotile) from meat
products, MLG 5.03, rev. 10/25/02. USDA/FSIS Microbiology Laboratory Guidebook, 3rd ed. U.S. Department of Agriculture, Food Safety Inspection Service, Washington, D.C.

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5/1/05
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