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BS EN ISO 21872-1:2017+A1:2023

BS EN ISO 21872-1:2017+A1:2023

$189.07

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 2023 46
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PDF Catalog

PDF Pages PDF Title
2 undefined
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 Blank Page
32 Blank Page
33 Blank Page
34 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
38 Annex D (informative) Real-time PCR for the detection of Vibrio parahaemolyticus, thermostable direct haemolysin gene (tdh) and Vibrio vulnificus
40 Annex E (informative) Results of an interlaboratory study
43 Bibliography

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BS EN ISO 21872-1:2017+A1:2023
$189.07