ISO/TS 21872-2:2020 PDF

Name: ISO/TS 21872-2:2020 — Microbiology of the food chain — Horizontal method for the determination of Vibrio spp. — Part 2: Enumeration of total and potentially enteropathogenic Vibrio parahaemolyticus in seafood using nucleic acid hybridization
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Microbiology of the food chain — Horizontal method for the determination of Vibrio spp. — Part 2: Enumeration of total and potentially enteropathogenic Vibrio parahaemolyticus in seafood using nucleic acid hybridization

Название (английский):

ISO/TS 21872-2:2020

Статус документа:: Действующий
Формат:: Электронный (PDF)
Количество страниц:: 27
Дата публикации:: 27 октября 2020 г.
Издание:: ISO TS 21872 edition 1 version 1
ICS:: 07.100.30

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Описание (английский):

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.

Abstract

Похожие стандарты

Стандарты, упомянутые в описании

Overview

ISO/TS 21872-2:2020 - "Microbiology of the food chain - Horizontal method for the determination of Vibrio spp. - Part 2" specifies a laboratory method for the direct enumeration of total Vibrio parahaemolyticus (toxR gene) and potentially enteropathogenic V. parahaemolyticus (tdh and/or trh positive) in seafood using nucleic acid hybridization. The technique combines traditional plating with nylon membrane transfer and hybridization using digoxigenin‑labelled oligoprobes, followed by colorimetric detection to enumerate target colonies.

Key topics and technical requirements

Purpose: Direct enumeration of total and potentially enteropathogenic V. parahaemolyticus in seafood samples.

Core principle: Plate appropriate dilutions on non‑selective nutrient medium → transfer colonies to membranes → lyse cells and cross‑link DNA → hybridize with labelled oligoprobes (tdh, trh, toxR) → colorimetric detection and colony counting.

Probes and targets: Digoxigenin‑labelled oligoprobes for tdh, trh (enteropathogenic markers) and toxR (species marker). Annex C contains probe and buffer information.

Controls: Process control, hybridization control and detection control are required to validate each stage.

Culture media & reagents: Composition and preparation specified (see Annex B); commercial reagents allowed if conformant. Follow ISO 11133 for media production and performance testing.

Recommended equipment: rotary blender, pipettes, incubator (37 °C ±1 °C), UV cross‑linker or transilluminator, hybridization oven/water bath, rocking platform, washing containers.

Procedure steps: Sampling and sample preparation (per ISO 6887 series), plating, colony lift, DNA cross‑linking, proteinase K treatment (if applicable), hybridization, colorimetric detection, optional isolation of positive colonies for confirmation.

Interpretation & reporting: Enumeration results, detection of tdh/trh and toxR positives, and test report elements are defined in the specification.

Applications and users

Who uses it: Food microbiology laboratories, seafood processors and quality control teams, public‑health laboratories, regulatory agencies, and research groups monitoring seafood safety.

Practical applications:

Routine surveillance of shellfish and other seafood for V. parahaemolyticus contamination.
Differentiation between total V. parahaemolyticus and potentially enteropathogenic strains (tdh/trh), improving risk assessment.
Supporting outbreak investigations by enumerating pathogenic subpopulations and guiding isolation for confirmatory testing.
Method standardization across labs to ensure consistent, comparable results for surveillance and compliance.

Related standards

ISO 7218 - General requirements for microbiological examinations

ISO 6887‑1 / ISO 6887‑3 - Sample preparation and dilutions for food and fish products

ISO 11133 - Preparation and performance testing of culture media

Keywords: ISO/TS 21872-2:2020, Vibrio parahaemolyticus, nucleic acid hybridization, seafood testing, tdh, trh, toxR, enumeration, microbiology of the food chain.

ISO/TS 21872-2:2020 PDF

Abstract

Похожие стандарты

Overview

Key topics and technical requirements

Applications and users

Related standards

Технические детали

ISO 11133:2014/Amd 2:2020

Microbiology of food, animal feed and water — Preparation, production, storage and performance testing of cul…

ISO 6887-3:2017/Amd 1:2020

Microbiology of the food chain — Preparation of test samples, initial suspension and decimal dilutions for mi…

SIST EN ISO 7218:2024

Microbiology of the food chain - General requirements and guidance for microbiological examinations (ISO 7218…

SIST EN ISO 6887-1:2017/A1:2024

Microbiology of the food chain - Preparation of test samples, initial suspension and decimal dilutions for mi…