BS EN ISO 21872-1:2017
$167.15
Microbiology of the food chain. Horizontal method for the determination of Vibrio spp. – Detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus
Published By | Publication Date | Number of Pages |
BSI | 2017 | 42 |
This document specifies a horizontal method for the detection of enteropathogenic Vibrio spp., which causes human illness in or via the intestinal tract. The species detectable by the methods specified include Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus.
It is applicable to the following:
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products intended for human consumption and the feeding of animals;
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environmental samples in the area of food production and food handling.
This method may not be appropriate in every detail for certain products (see Introduction).
The World Health Organization (WHO) has identified that V. parahaemolyticus, V. cholerae and V. vulnificus are the major food-borne Vibrio spp. However, the method in this document can also be appropriate for the identification of other Vibrio spp. causing illness in humans. [1]
PDF Catalog
PDF Pages | PDF Title |
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2 | National foreword |
4 | European foreword Endorsement notice |
7 | Foreword |
8 | Introduction |
9 | 1 Scope 2 Normative references |
10 | 3 Terms and definitions 4 Principle 4.1 General 4.2 Primary enrichment in a liquid selective medium |
11 | 4.3 Secondary enrichment in a liquid selective medium 4.4 Isolation and identification 4.5 Confirmation 5 Culture media and reagents |
12 | 5.1 Enrichment medium: alkaline saline peptone water (ASPW) 5.2 Solid selective isolation media 5.2.1 First medium: thiosulphate, citrate, bile and sucrose agar medium (TCBS) 5.2.2 Second medium 5.3 Saline nutrient agar (SNA) 5.4 Reagent for detection of oxidase 5.5 Biochemical tests 5.5.1 L-lysine decarboxylase saline medium (LDC) 5.5.2 Arginine dihydrolase saline medium (ADH) |
13 | 5.5.3 Reagent for detection of β-galactosidase 5.5.4 Saline medium for detection of indole 5.5.5 Saline peptone waters 5.5.6 Sodium chloride solution 5.6 PCR 5.6.1 Tris acetate EDTA buffer (TAE) (or a buffer allowing similar performance for the purpose) 5.6.2 Mastermix 5.6.3 Primers and probes 5.6.4 Positive control material |
14 | 5.6.5 Negative extraction control 6 Equipment and consumables 7 Sampling 8 Preparation of the test sample |
15 | 9 Procedure (See Figure A.1) 9.1 Test portion and initial suspension 9.2 Primary selective enrichment 9.3 Secondary selective enrichment |
16 | 9.4 Isolation and identification 9.5 Confirmation 9.5.1 General |
17 | 9.5.2 Selection of colonies for confirmation and preparation of pure cultures 9.5.3 Tests for presumptive identification |
18 | 9.5.4 Biochemical confirmation |
20 | 9.5.5 PCR confirmation 9.5.6 DNA extraction 9.5.7 Conventional PCR |
21 | 9.5.8 Real-time PCR 10 Expression of results 11 Performance characteristics of the method 11.1 Interlaboratory study |
22 | 11.2 Sensitivity 11.3 Specificity 11.4 LOD50 12 Test report |
23 | Annex A (normative) Diagram of procedure |
25 | Annex B (normative) Composition and preparation of the culture media and reagents |
31 | Annex C (informative) Conventional PCR for the detection of Vibrio parahaemolyticus, thermostable direct haemolysin (tdh) and thermostable direct related haemolysin (trh) genes, Vibrio cholerae and Vibrio vulnificus |
35 | Annex D (informative) Real-time PCR for the detection of Vibrio parahaemolyticus, thermostable direct haemolysin gene (tdh) and Vibrio vulnificus |
37 | Annex E (informative) Results of an interlaboratory study |
40 | Bibliography |