MFLP-96 The Reveal Test Kit for Detecting Salmonella
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193922F26A0648868654BDA1C0109B00 |
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0.02 |
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页数： |
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-96,February 2001,THE REVEAL TEST KIT FOR DETECTING SALMONELLA,1. APPLICATION,The method is applicable to the detection of Salmonella from feeds, raw foods and other samples to,determine compliance with Sections 4 and 7 of the Food and Drugs Act. Where stipulated, the Official,Methods and HPB Method should be used.,2. DESCRIPTION,The method has been shown to produce satisfactory results with artificially contaminated and naturally,contaminated foods in AFNOR studies (8.1).,3. PRINCIPLE,Following selective enrichment in Rappaport-Vasiliadis broth, a portion of the sample enrichment is placed,into the sample port of the Reveal Device initiating flow. The Reveal Device contains antibodies with high,specificity to Salmonella antigens. These antibodies are bound to colloidal gold and, separately, to a solid,support matrix. Any Salmonella antigen present will bind to the gold conjugated antibodies forming an,antigen – antibody – chromogen complex. This complex flows across a lateral flow membrane and is,subsequently bound by antibody immobilized on the membrane. This causes the gold conjugate to,precipitate, forming a visible line and indicating a positive reaction. Proper test completion and flow is,indicated by a control line which forms further up in the test window and verifies a valid test run. Absence,of a control line invalidates the test. Record results at 10 minutes incubation time.,4. DEFINITION OF TERMS,See Appendix A of Volume 3.,5. COLLECTION OF SAMPLES,See Appendix B of Volume 3.,6. MATERIALS AND SPECIAL EQUIPMENT,The media listed under No. 2 to 7 are commercially available and are to be prepared and sterilized,according to the manufacturer’s instructions.,1) Reveal Device (Neogen Corporation, phone: (517) 372-9200, Fax: (517) 372-0108),2) REVIVE broth (Neogen Corp., Lansing, MI),3) Rappaport-Vasiliadis broth at 2X concentration (Neogen Corp., Lansing, MI),MFLP-96,- 2 - February 2001,4) Xylose lysine desoxycholate agar (XLD),5) Hektoen enteric agar (HE),6) Bismuth sulfite agar (BS),7) Brilliant green sulfa agar (BGS),8) Incubator capable of maintaining 35EC.,9) Incubator capable of maintaining 42 EC.,NOTE: It is the responsibility of each laboratory to ensure that the temperature of the incubators or water baths are,maintained at the recommended temperatures. Where 35EC is recommended in text of the method the,incubator may be at 35 +/-1.0E C. Similarly, lower temperatures of 30 or 25 may be +/- 1.0EC. However,where higher temperatures are recommended, such as 43 or 45.5EC, it is imperative that the incubators or,water baths be maintained within 0.5EC due to potential lethality of higher temperatures on the,microorganism being isolated.,11) Stomacher, blender or equivalent,12) Stomacher bags and screw-capped jars,13) Micropipetter and sterile tips,14) Control cultures (ATCC or equivalent),15) pH paper,16) 1N NaOH and 1N Hcl,17) Biochemicals and antisera listed in MFHPB-20,7. PROCEDURE,Each sample unit may be analyzed individually or the analytical units may be combined. Carry out the test,in accordance with the following instructions:,7.1 Handling of Sample Units,7.1.1 In the laboratory prior to analysis, keep sample units refrigerated (0-5EC) or frozen,depending on the nature of the product. Thaw frozen samples in a refrigerator, or under,time and temperature conditions which prevent microbial growth or death.,7.1.2 Analyze sample units as soon as possible after their receipt in the laboratory,7.2 Preparation for Analysis,7.2.1 Have ready REVIVE broth (see Table 1).,7.2.2 Clean the surface of the working area with a suitable disinfectant,MFLP-96,- 3 - February 2001,7.3 Preparation of Sample,7.3.1 To ensure a truly representative analytical unit agitate liquids or free flowing materials until,the contents are homogeneous. If a sample unit is a solid, obtain the analytical unit by,taking a portion from several locations within the sample unit. To reduce the workload, the,analytical units may be combined for analysis. It is recommended that a composite contain,not more than 500g.,7.3.2 Prepare a 1:9 dilution of the food by aseptically blending or stomaching 25 g or mL (the,analytical unit) into 200 mL REVIVE, as indicated in Table 1.,7.3.3 Adjust pH of the mixture, if necessary, to 6.8 ± 0.2 with 1 N NaOH or 1 N HCl.,7.3.4 Incubate preenrichment mixture 4 h at 35EC.,7.4 Selective Enrichment of Sample,Gently shake incubated sample mixture and add 200 mL of 2X Rappaport-Vasiliadis broth (RV).,Incubate ……
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