ISO/TS 20224-1:2020 PDF

Molecular biomarker analysis — Detection of animal-derived materials in foodstuffs and feedstuffs by real-time PCR — Part 1: Bovine DNA detection method

Название (английский):

ISO/TS 20224-1:2020

Molecular biomarker analysis — Detection of animal-derived materials in foodstuffs and feedstuffs by real-time PCR — Part 1: Bovine DNA detection method

Статус документа:: Действующий
Формат:: Электронный (PDF)
Количество страниц:: 16
Дата публикации:: 31 июля 2020 г.
Издание:: ISO TS 20224 edition 1 version 1
ICS:: 67.050

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Описание (английский):

This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of bovine-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of bovine material derived from cattle including taurine (Bos taurus) and zebu (Bos indicus). The assay also detects the species bison (Bison bison) and yak (Bos mutus). The target sequence is a partial fragment of the bovine nuclear beta actin gene in chromosome 25 (i.e. GenBank accession number NC_037352.1)[1][2][3], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

Технические детали

Технический комитет: ISO/TC 34/SC 16 - Horizontal methods for molecular biomarker analysis
SKU: ISO/TS 20224-1:2020

Abstract

Overview

ISO/TS 20224-1:2020 specifies a real-time PCR method for qualitative detection of bovine DNA in foodstuffs and feedstuffs. The technique targets a partial fragment of the bovine nuclear beta actin gene on chromosome 25 (GenBank NC_037352.1), a single-copy locus per haploid genome. The assay is designed for routine bovine DNA detection (Bos taurus and Bos indicus) and also detects bison (Bison bison) and yak (Bos mutus). The provided PCR assay has an absolute limit of detection of five copies per reaction with ≥95% replicability (LOD95%).

Key topics and technical requirements

Scope: Qualitative species identification of bovine-derived material in food and feed using real-time PCR.
Target: Partial fragment of bovine nuclear beta actin gene (NC_037352.1), single-copy per haploid genome.
Sensitivity: Absolute LOD = 5 copies/reaction, ≥95% replicability at LOD95%.
Sample preparation: Requires extraction of an adequate amount of PCR-amplifiable DNA from the test matrix following ISO 21571 recommendations.
Reaction setup (typical):
- Total volume: 25 µl; sample DNA 5 µl (recommended 20–200 ng/µl).
- 2× PCR master mix 12.5 µl; primers and probe at specified concentrations (primers ~400 nmol/L, probe ~200 nmol/L).
- Compatible with TaqMan-style real-time PCR chemistry; master mix may include UDG/dUTP to limit carry-over contamination.
Controls and verification:
- Include inhibition/reference gene assay (e.g., 18S rRNA or myostatin) to confirm DNA amplifiability and avoid false negatives.
- Standard PCR controls and plate layout guidance to mitigate edge effects and contamination.
Apparatus: Real-time thermocycler capable of fluorescence excitation/detection for TaqMan assays.

Applications and users

ISO/TS 20224-1:2020 is intended for laboratories performing:

Food authenticity and fraud detection (identifying undeclared bovine ingredients).
Regulatory and enforcement testing for food labeling and feed compliance.
Supply-chain verification for religious (halal/kosher) or cultural dietary requirements.
Quality control in food manufacturers, feed producers, and testing service providers. Typical users: public health and food control laboratories, contract testing labs, industry QC teams, and forensic analysts focused on species identification.

Related standards

ISO 21571 - nucleic acid extraction for foodstuffs (recommended extraction methods)
ISO 20813 - general requirements for nucleic acid-based species detection
ISO 24276 - PCR method controls and requirements
ISO 16577 - molecular biomarker analysis terminology

Keywords: ISO/TS 20224-1:2020, bovine DNA detection, real-time PCR, beta actin gene, food authenticity, species identification, LOD95%, food and feed testing.

Overview

Key topics and technical requirements

Scope: Qualitative species identification of bovine-derived material in food and feed using real-time PCR.
Target: Partial fragment of bovine nuclear beta actin gene (NC_037352.1), single-copy per haploid genome.
Sensitivity: Absolute LOD = 5 copies/reaction, ≥95% replicability at LOD95%.
Sample preparation: Requires extraction of an adequate amount of PCR-amplifiable DNA from the test matrix following ISO 21571 recommendations.
Reaction setup (typical):
- Total volume: 25 µl; sample DNA 5 µl (recommended 20–200 ng/µl).
- 2× PCR master mix 12.5 µl; primers and probe at specified concentrations (primers ~400 nmol/L, probe ~200 nmol/L).
- Compatible with TaqMan-style real-time PCR chemistry; master mix may include UDG/dUTP to limit carry-over contamination.
Controls and verification:
- Include inhibition/reference gene assay (e.g., 18S rRNA or myostatin) to confirm DNA amplifiability and avoid false negatives.
- Standard PCR controls and plate layout guidance to mitigate edge effects and contamination.
Apparatus: Real-time thermocycler capable of fluorescence excitation/detection for TaqMan assays.

Applications and users

ISO/TS 20224-1:2020 is intended for laboratories performing:

Food authenticity and fraud detection (identifying undeclared bovine ingredients).
Regulatory and enforcement testing for food labeling and feed compliance.
Supply-chain verification for religious (halal/kosher) or cultural dietary requirements.
Quality control in food manufacturers, feed producers, and testing service providers. Typical users: public health and food control laboratories, contract testing labs, industry QC teams, and forensic analysts focused on species identification.

Related standards

ISO 21571 - nucleic acid extraction for foodstuffs (recommended extraction methods)
ISO 20813 - general requirements for nucleic acid-based species detection
ISO 24276 - PCR method controls and requirements
ISO 16577 - molecular biomarker analysis terminology

Keywords: ISO/TS 20224-1:2020, bovine DNA detection, real-time PCR, beta actin gene, food authenticity, species identification, LOD95%, food and feed testing.

ISO/TS 20224-1:2020 PDF

Технические детали

Abstract

Overview

Key topics and technical requirements

Applications and users

Related standards

Похожие стандарты

Abstract

Overview

Key topics and technical requirements

Applications and users

Related standards

Похожие стандарты

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