MFHPB-30 Isolation of Listeria monocytogenes from All Foods and Environmental Samples
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E7E13714488446FB87DA348DF19960B1 |
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文件大小(MB)： |
0.1 |
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页数： |
13 |
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日期： |
2012-3-2 |
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Published on the Food Directorate’s (Health Canada's) website at:,http://www.hc-sc.gc.ca/fn-an/res-rech/analy-meth/microbio/index-eng.php,Government of Canada Gouvernement du Canada,HPB Method MFHPB-30,February 2011,Health Products and Food Branch,Ottawa,Isolation of Listeria monocytogenes and other Listeria spp.,from foods and environmental samples,Franco Pagotto, Karine Hébert and Jeff Farber,Bureau of Microbial Hazards,Food Directorate,Health Canada,Postal locator: 2204E,Ottawa, ON, KlA 0K9,Email: micro_methods_committee@hc-sc.gc.ca,1. Application,This method is applicable to the isolation and identification of Listeria monocytogenes and other Listeria,species to determine compliance with the requirements of Sections 4 and 7 of the Food and Drugs Act. This,method is Health Canada’s standard reference cultural method and can be used for all foods and,environmental surfaces. This revised method replaces MFHPB-30, dated September 2010, the Supplement to,MFHPB-30, dated March 2002, and Appendix L (Supplement to All Listeria Methods), dated August 2005.,2. Principle,This method determines the presence of viable Listeria species in food and environmental samples. A portion,of the product (or environmental sample such as a sponge or swab) is enriched first in a primary broth, then in,a secondary enrichment broth, plated onto a specified agar medium and one additional plating medium, and,then incubated under specified conditions of time and temperature. It is assumed that viable Listeria cells will,multiply under these conditions and give rise to visible colonies which can be identified. This method is based,on those of Lovett (8.4, 8.5), Hitchins (8.3) and McClain and Lee (8.6), that have been modified on the basis of,data collected in comparative studies by Warburton et al. (8.13, 8.14, 8.15). It was revised by the addition of,modified Fraser broth (8.7), lithium chloride-phenylethanol-moxalactam medium (8.6), and Oxford (8.13),modified Oxford (8.7), and/or PALCAM (8.15) agars. Further revisions to include other chromogenic media,have also been made. A mandatory step has been added that requires 24 h enriched LEB (UVM1 formulation),broth to be directly streaked to selective agars, in addition to a transfer step to modified Fraser broth.,- 2 -,MFHPB-30,February 2011,Published on the Food Directorate’s (Health Canada's) website at:,http://www.hc-sc.gc.ca/fn-an/res-rech/analy-meth/microbio/index-eng.php,3. Definition of terms,See Appendix A of Volume 2.,4. Collection of samples,4.1 See Appendix B of Volume 2.,4.2 For environmental samples, refer to the sampling procedures given in MFLP-41.,5. Materials and special equipment,Note: The Laboratory Supervisor must ensure that the analysis described in this method is carried out in,accordance with the International Standard referred to as “ISO/IEC 17025:2005 (or latest,version): General Requirements for the Competence of Testing and Calibration Laboratories".,The media and reagents listed below that are commercially available and are to be used, prepared and/or,sterilized according to the manufacturer's instructions. See Appendix G of Volume 2 for the media,formula.,Note: If the analyst uses any variations of the media listed here (either product that is commercially,available or made from scratch), it is the responsibility of the analyst or Laboratory Supervisor to,ensure equivalency. Please forward equivalency data to the Editor of Compendium of,Analytical Methods for consideration of modification of this method.,Listeria broths and agars (base media and supplements are commercially available),1) Listeria enrichment broth (LEB) - UVM1 formulation,2) Modified Fraser broth (MFB),3) Oxford agar (OXA) - mandatory plating media,4) Plating media for second selective agar (one of the following is mandatory),- Agar Listeria according to Ottaviana and Agosti (ALOA),- A.L. Agar (Bio-Rad),- BBL CHROMagar Listeria (BD),- Chromogeneic Listeria Agar Plate (OCLA; Oxoid),- Lithium chloride-phenylethanol-moxalactam medium (LPM),- Modified Oxford agar (MOX),- PALCAM agar (PAL),- RAPID’L.Mono (Bio-Rad),5) Control cultures (use ATCC strains or equivalent),Positive controls: Listeria monocytogenes, Listeria ivanovii, Listeria innocua, Listeria grayi,(Staphylococcus aureus and Rhodococcus equi - optional),6) Stomacher, blender or equivalent, vortex mixer,7) Microscope,- 3 -,MFHPB-30,February 2011,Published on the Food Directorate’s (Health Canada's) website at:,http://www.hc-sc.gc.ca/fn-an/res-rech/analy-meth/microbio/index-eng.php,8) Incubators capable of maintaining 30oC and 35oC,Note: It is the responsibility of each laboratory to ensure that the temp……
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