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BS EN ISO 21872-1:2017+A1:2023 Microbiology of the food chain. Horizontal method for the determination of Vibrio spp. - Detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus >
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BS EN ISO 21872-1:2017+A1:2023 Microbiology of the food chain. Horizontal method for the determination of Vibrio spp. - Detection of potentially enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus, 2023
- undefined
- European foreword
- Endorsement notice
- Foreword
- Introduction
- 1 Scope
- 2 Normative references
- 3 Terms and definitions
- 4 Principle
- 4.1 General
- 4.2 Primary enrichment in a liquid selective medium
- 4.3 Secondary enrichment in a liquid selective medium
- 4.4 Isolation and identification
- 4.5 Confirmation
- 5 Culture media and reagents
- 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)
- 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
- 5.6.5 Negative extraction control
- 6 Equipment and consumables
- 7 Sampling
- 8 Preparation of the test sample
- 9 Procedure (See Figure A.1)
- 9.1 Test portion and initial suspension
- 9.2 Primary selective enrichment
- 9.3 Secondary selective enrichment
- 9.4 Isolation and identification
- 9.5 Confirmation
- 9.5.1 General
- 9.5.2 Selection of colonies for confirmation and preparation of pure cultures
- 9.5.3 Tests for presumptive identification
- 9.5.4 Biochemical confirmation
- 9.5.5 PCR confirmation
- 9.5.6 DNA extraction
- 9.5.7 Conventional PCR
- 9.5.8 Real-time PCR
- 10 Expression of results
- 11 Performance characteristics of the method
- 11.1 Interlaboratory study
- 11.2 Sensitivity
- 11.3 Specificity
- 11.4 LOD50
- 12 Test report
- Annex A (normative) Diagram of procedure
- Annex B (normative) Composition and preparation of the culture media and reagents
- Blank Page
- Blank Page
- Blank Page
- 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
- Annex D (informative) Real-time PCR for the detection of Vibrio parahaemolyticus, thermostable direct haemolysin gene (tdh) and Vibrio vulnificus
- Annex E (informative) Results of an interlaboratory study
- Bibliography [Go to Page]