Annex D-1

Methodology for Escherichia coli O157:H7 testing of beef products

As of July 26, 2011 the CFIA currently has equivalency with the United States - FSIS for the following screening and cultural methods for E. coli O157:H7, Salmonella spp. and Listeria monocytogenes:

E. coli O157:H7 in Meat

1. MFLP-16, Detection of E. coli O157:H7 in Foods - Assurance GDS^TM for E. coli O157:H7 Gene Detection System, August 2005, with the following stipulations:
   • testing of a 375 ± 37 gram test portion, analyzed as a 1:3.2 dilution in pre-warmed modified EnteroHaemorrhagic Escherichia Coli (mEHEC) broth.
   • incubated for 10 to 18 hours at 42°C
2. MFLP-16, Detection of E. coli O157:H7 in Foods - Assurance GDS^TM for E. coli O157:H7 Gene Detection System, August 2005, with the following stipulations:
   • testing of a 325 ± 32 gram test portion, analyzed as a 1:9 dilution in pre-warmed mEHEC media
   • incubated for 10 to 18 hours at 42°C
3. MFLP-30, The DuPont Qualicon Bax® System Method for the Detection of E. coli O157:H7 in Raw Beef and Fruit Juice, May 2003, including both Supplement 1 (May 2005) and Supplement 2 (November 2006), with the following stipulation:
   • testing of a 325 ± 32 gram test portion
4. MFLP-87, Detection of Enterohemorrhagic E. coli (EHEC) in Food Products and Food Ingredients by the VIP for EHEC Method, June 2005, with the following stipulation:
   • testing of a 325 ± 32 gram test portion
5. MFLP-19, The DuPont Lateral Flow System^TM Method for Detecting E. coli O157:H7 in Raw Ground and Raw Boneless Beef, March 2006, including Supplement 1, August 2007, with the following stipulation:
   • testing of a 325 ± 32 gram test portion
6. MFLP-12, Identification of E. coli O157:H7 and Verotoxin-producing E. coli O157 Non Motile (NM) by the Warnex^TM Real-time Polymerase Chain Reaction System, January 2006, with the following stipulation:
   • testing of a 325 ± 32 gram test portion
7. MFLP-80, Isolation of E. coli O157:H7 or NM in Foods, March 2008 (cultural confirmation method), including MFLP-90 (IMS isolation method) and MFLP-83 or MFLP-93 (verotoxin confirmation methods) referred to in MFLP-80, with the following stipulations:
   • testing of a 325 gram test portion
   • typical E. coli O157:H7 colonies on selective agar that do not confirm as a toxin producer must be submitted to CFIA for further PCR testing of toxin genes and H7 genes
8. MFLP-76, The DuPont Qualicon BAX ®System real-time method for the detection of E. coli 0157:H7, 2011, in raw beef trim and ground beef, with the following stipulations:
   • testing of a 325 gram test portion

Salmonella spp. in Meat and Eggs

1. MFLP-29, The Qualicon Bax® System Method for the Detection of Salmonella in a Variety of Food and Environmental Samples, July 2007, with the following stipulations:
   • for meat products, analyse a 325 gram test portion
   • for egg products, analyse a 250 gram test portion*
2. MFHPB-24, Detection of Salmonella spp. in Foods by the Vidas SLMTM Method^TM, November 2001, with the following stipulations:
   • for meat products, analyse a 325 gram test portion
   • for egg products, analyse a 250 gram test portion*
3. MFHPB-20, Isolation and Identification of Salmonella from Foods, April 1998 (cultural confirmation method), with the following stipulations:
   • for meat products, analyse a 325 gram test portion
   • for egg products, analyse a 250 gram test portion*

* USDA/FSIS requirements for egg products exported to the U.S. are for analysis of a 100 gram test portion.

Salmonella spp. in Meat

1. MFLP-32, Identification of Salmonella species by the Warnex^TM Real Time Polymerase Chain Reaction System, June 2005, with the following stipulations:
   • for meat products, analyse a 325 gram test portion

Listeria monocytogenes in Meat and Eggs

1. MFLP-28: The Qualicon Bax® System Method for the Detection of Listeria monocytogenes in a Variety of Food, June 2003, and the Supplement to the method, January 2004, with the following stipulations:
   • for meat and egg products, analyse a 25 gram test portion
2. MFHPB-30: Isolation of Listeria monocytogenes from all Food and Environmental Samples, January 2001 (cultural confirmation method), and the Supplement to the method, March 2002, with the following stipulations:
   • for meat and egg products, analyse a 25 gram test portion
3. MFLP-15: The Determination of Listeria spp. from Environmental Surfaces Using The DuPont Qualicon Bax® System Method and Direct Plating, February 2009, with the following stipulations:
   • swabs of environmental surfaces may be tested individually or composited (up to 10 swabs)

In addition FSIS will accept all methods for the above pathogens published in the USDA/FSIS Microbiology Laboratory Guidebook.

Please direct questions or comments to one of the following contacts:

Donna Douey
Office: 403-299-7686
Cell: 403-836-2642
e-mail: Donna.Douey@inspection.gc.ca

Sam Mohajer
Office: 613-773-5307
Cell: 613 371 0423
e-mail: Sam.Mohajer@inspection.gc.ca

Johanna Murphy
Office: 613-773-5413
Cell: 613-327-0126
e-mail: Johanna.Murphy@inspection.gc.ca

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