MFLP-35 Detection of Salmonella by the Tecra Salmonella Visual Immunoassay (Via) Method
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FDAFB03D379146C68DCA9FF9E9B6E059 |
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0.09 |
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5 |
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日期： |
2024-7-30 |
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Published on the Food Directorate’s (Health Canada's) website at http://www.hc-sc.gc.ca/food-aliment.,Government of Canada Gouvernement du Canada,Laboratory Procedure MFLP-35,June 2002,HEALTH PRODUCTS AND FOOD BRANCH,OTTAWA,DETECTION OF SALMONELLA BY THE TECRAa,SALMONELLA VISUAL IMMUNOASSAYTM (VIA) METHOD,Evaluation Division,Bureau of Microbial Hazards,Food Directorate, Health Canada,Postal locator: 2204A1,Ottawa, Ontario, K1A 0L2,E-mail: don_warburton@hc-sc.gc.ca,1. APPLICATION,This method is applicable to the detection of Salmonella from foods, food ingredients and,environmental samples to determine compliance with the requirements of Sections 4 and 7 of the,Food and Drugs Act.,The TECRA Salmonella Visual Immunoassay is an Enzyme-linked Immunosorbent assay (ELISA),which provides a rapid and specific screening system for the in vitro detection of Salmonella in food,and environmental samples, after selective enrichment. Results are determined either visually or,using a microtitre plate reader.,Presumptive positive results should be confirmed by standard cultural procedures. This is especially,important in situations such as product recalls.,The TECRA Salmonella VIA has AOAC Official Methods approval for use with several different,enrichment broths (AOAC methods 989.14 and 998.09). It also has approvals in several countries,including France, Australia, New Zealand, Denmark and Brazil.,1. PRINCIPLE,The TECRA Salmonella Visual immunoassay is an Enzyme linked Immunoasorbent Assay,(ELISA) performed in the “sandwich” configuration. High affinity “capture” antibodies specific for,Salmonella have been adsorbed onto the surface of wells. If Salmonella antigens are present in,the sample, they are captured by the antibodies. All other material in the sample is washed away.,The “sandwich” is completed by the addition of enzyme labelled antibodies (conjugate) specific for,Salmonella. The presence of Salmonella is indicated when the bound conjugate converts the,substrate to a green color. Alternatively, if Salmonella is not present, no green color develops.,Total time to perform the test including selective enrichment and post enrichment in M broth is 42-,50 hours.,MFLP-35,- 2 - June 2002,If a presumptive positive Salmonella result is obtained with the VIA, this should be confirmed by,streaking from the enrichment broth onto selective agar plates.,TECRAa is the registered trademark of TECRA International Pty Ltd.13 Rodborough Rd, Frenchs,Forest, NSW 2086 Australia.,3. DEFINITIONS OF TERMS,See Appendix A of Volume 3.,3.1 Sampling,See Appendix B of Volume 3.,3.2 Stringency of Sampling,See MFHPB-20 and implement if applicable.,3.3 “Routine” versus “Investigational” Sampling Plans,See MFHPB-20 and implement if applicable.,4. MATERIALS AND SPECIAL EQUIPMENT,4.1 Kit Components :,All reagents for the TECRA Salmonella Visual Immunoassay are provided in one kit, which contains,enough material for a maximum of 94 tests (96 wells are provided but the positive and negative,controls supplied with the kit must be run each time the test is performed). The kits are available in,North America from International Bioproducts Inc. 21312 30th Drive SE, Bothell WA 98021.,Instructions, sample record sheet,Plate holder,Resealing strip,Antibody coated microtitre wells,Reagent 1 - Wash concentrate,Reagent 2 - Positive control,Reagent 3 - Control diluent,Reagent 4 - Conjugate,Reagent 5 - Conjugate diluent,Reagent 6 - Substrate,Reagent 7 - Substrate diluent,Reagent 8 - Stop Solution,4.2 Additional Requirements :,35-37/C incubator,41-43/C incubator,Serological pipettes,Plastic squeeze bottle,Pipettes (20:L, 50 :L, and 200 :L),Plastic wrap,Waterbath (100/C),Small screw cap bottles or tubes capable of containing 5 mL and suitable for boiling.,Vortex mixer,MFLP-35,- 3 - June 2002,Glass test tubes (15cm x 15cm),Blender/stomacher/stomacher bags.,4.3 Media (VIA),Media are commercially available and are to be prepared and sterilized according to the,manufacturer's instructions. See also Appendix G of Volume 3 and reference 8.2 for the formula of,individual media.,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.,Enrichment broth media:,Different enrichment protocols are described in the TECRA Methods Manual included in each kit.,Protocol 1: AOAC method 998.09 Option 1 RV(R10)/TT,Lactose broth or other non inhibitory broth as specified in FDA BAM (pre-enrichment),Tetrathionate broth and Rappaport Vassiliadis (RV-R10) broth (selective enrichment),M broth……
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