MFLP-72 The Isolation and Identification of Vibrio cholerae 01 and non-01 from Foods
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BA5C9973B6D445829F1D1F3100414486 |
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文件大小(MB)： |
0.06 |
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页数： |
16 |
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日期： |
2012-3-2 |
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Published by: POLYSCIENCE PUBLICATIONS, P.O. Box 1606, Station St-Martin, Laval, Quebec,Canada H7V 3P9. TEL.: 1-800-840-5870. FAX: (450) 688-1930.,Government of Canada Gouvernement du Canada,Laboratory Procedure MFLP-72,April 1995,HEALTH PROTECTION BRANCH,OTTAWA,THE ISOLATION AND IDENTIFICATION OF,Vibrio cholerae O1 AND NON-O1 FROM FOODS,S. Stavric and B. Buchanan,Research Division, Bureau of Microbial Hazards, Food Directorate,Health Canada, Ottawa ON, K1A 0L2,1. APPLICATION,This method is applicable to the detection of Vibrio cholerae O1 and non-O1 in foods to determine,compliance with the requirements of Sections 4 and 7 of the Food and Drugs Act.,2. DESCRIPTION,This method has been shown to produce satisfactory results with fish, seafood, vegetables and coconut,milk. V. cholerae classification is based upon agglutination in antisera directed against the somatic "O",antigens. Species that agglutinate in anti-O1 serum form serogroup O1, while those that do not, form,serogroup non-O1 (9.3). Both O1 and non-O1 serogroups include strains that produce cholera toxin.,Serogroup O1 is further divided into two biotypes, Classical and El Tor, based upon certain physiological,and bacteriophage sensitivity patterns; and three serotypes, Inaba, Ogawa, or Hikojima, based on,agglutination with specific antisera (9.5, 9.7). Strains isolated from the 1992/93 epidemic in India and,Bangladesh were assigned a new serogroup O139, with a synonym "Bengal" (9.8). They can be identified,by performing a battery of biochemical tests used for the 01 and non-01 serogroups, followed by the,serological confirmation with O139 antiserum (9.6).,3. PRINCIPLE,Detection of this organism requires three successive phases: (i) enrichment in a nonselective medium, (ii),plating out on a selective medium and presumptive identification, and (iii) confirmation by biochemical and,serological tests. This method is based on that of Elliot et al. published recently in the Bacteriological,Analytical Manual of the United States Food and Drug Administration (9.2). It is recommended that all,isolates from serogroups 01 and 0139 be tested for production of cholera toxin.,4. DEFINITION OF TERMS,See Appendix A of Volume 3.,5. COLLECTION OF SAMPLES,5.1 See Appendix B of Volume 3.,5.2 If the analysis is not performed immediately upon the arrival of samples in the laboratory, store,samples in the refrigerator (0-4EC) or frozen, depending on the nature of the product.,MFLP-72,- 2 - April 1995,6. MATERIALS AND SPECIAL EQUIPMENT,1) Alkaline peptone water (APW),2) Thiosulfate-citrate-bile salts-sucrose agar (TCBS),3) Tryptone salt agar (T1N1),4) Trypticase soy agar (TSA),5) Triple sugar iron agar (TSI),6) Kligler iron agar (KIA),7) Arginine glucose slants (AGS),8) Gelatin agar (GA),9) Gelatin salt agar (GSA),10) Hugh-Leifson glucose broth,11) Oxidase reagent,12) o-Nitrophenyl-$-D-galactosidase (ONPG) reagent,13) Antiserum for group non-O1 serotype O139. (Available from Dr. W. Johnson, National Laboratory,for Enteric Pathogens, LCDC, Health Canada, Ottawa, Tel. No. (613) 957-1386),14) Diagnostic antisera for group O1 and subgroups Inaba, Ogawa and Hikojima (Available from,Wellmark Diagnostics Ltd., Guelph, Ontario),15) Brain heart infusion agar (BHIA),16) Heart infusion agar (HIA),17) Mueller-Hinton agar,18) Heart infusion broth (HIB),19) Polymyxin B solution,20) Sheep red blood cells - 5%,21) Blood agar,22) Methyl red-Voges Proskauer broth (MR-VP) and reagents,23) Chicken red blood cells - 2.5%,24) Falkow's decarboxylase basal medium, with or without individually added amino acids e.g. lysine,and arginine,25) Trypticase soy broth (TSB),26) Tryptone (1%) broth with or without added NaCl to a final concentration of 0, 3, 6, 8 and 10% NaCl,MFLP-72,- 3 - April 1995,27) Bromcresol purple broths, with added sucrose or arabinose,28) O/129 (2,4-diamino-6,7-diisopropyl pteridine) - discs containing 10 and 150 μg,29) Waring blender, or equivalent,30) Incubators capable of maintaining 35±0.5EC and 42±0.5EC,7. PROCEDURE,Each sample unit may be analyzed individually or the analytical units may be composited. For a flow chart,of the complete analytical procedure, see Fig. 1, Schedule for Sample Analysis. 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 (0-4EC) or frozen, depending on,the nature of the product.,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 sterile APW.,7.3 Sample Analysis,7.3.1 The following guidelines apply to sample preparati……
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