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BSI PD ISO/TS 21872-2:2020

$167.15

Microbiology of the food chain. Horizontal method for the determination of Vibrio spp. – Enumeration of total and potentially enteropathogenic Vibrio parahaemolyticus in seafood using nucleic acid hybridization

Published By Publication Date Number of Pages
BSI 2020 36
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This document specifies a method for the direct enumeration of potentially enteropathogenic V. parahaemolyticus (tdh and/or trh positive) and/or the enumeration of total V. parahaemolyticus in seafood.

PDF Catalog

PDF Pages PDF Title
2 undefined
6 Foreword
7 Introduction
9 1 Scope
2 Normative references
3 Terms and definitions
10 4 Principle
4.1 General
4.2 Plating
4.3 Preparation of membranes
4.4 Hybridization
4.5 Detection
11 5 Culture media and reagents
5.1 General
5.2 Hybridization reagents
5.2.1 Hybridization buffer
5.2.2 Digoxigenin-labelled oligoprobes
5.2.3 Control materials
6 Equipment and consumables
12 7 Sampling
8 Procedure
8.1 General
8.2 Preparation of the sample
8.3 Plating
8.4 Preparation of process control
8.5 Preparation of membranes
8.5.1 Colony lift
13 8.5.2 Cross-linking of DNA to the membrane
8.5.3 Proteinase K treatment
8.6 Hybridization
8.6.1 General
14 8.6.2 Hybridization of tdh and trh probes
8.6.3 Colorimetric detection
15 8.6.4 Enumeration of tdh and/or trh positive colonies
8.6.5 Isolation of colonies (optional)
8.6.6 Hybridization of toxR probe
16 8.6.7 Colorimetric detection
8.6.8 Enumeration of toxR positive colonies
9 Interpretation of the results
17 10 Expression of results
11 Test report
18 Annex A (normative) Flow diagram for the enumeration of total and potentially enteropathogenic Vibrio parahaemolyticus in seafood using nucleic acid hybridization
21 Annex B (normative) Culture media and reagents
27 Annex C (informative) Hybridization of tdh, trh and toxR genes of V. parahaemolyticus
35 Bibliography
BSI PD ISO/TS 21872-2:2020
$167.15