ISO/TS 20224-7:2020 分子バイオマーカー分析—リアルタイムPCRによる食品および飼料中の動物由来物質の検出—パート7：ロバのDNA検出法 | ページ 2

Foreword

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This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 16, Horizontal methods for molecular biomarker analysis.

A list of all parts in the ISO 20224 series can be found on the ISO website.

Introduction

Fraudulent adulteration of meat in food and feed threatens both public safety and commerce. Adulteration can affect those adhering to ethnological dietary rules, economic development and social stability. This document provides a real-time polymerase chain reaction (real-time PCR) analytical method for the identification of meat animal species from nucleic acid present in the ingredients of food and feed.

Animal-derived biological materials in food and feed are detected and identified in the laboratory with the following successive (or simultaneous) steps: preparation of the test portion/sample, nucleic acid extraction and purification, PCR amplification and interpretation of results. This document provides guidance for PCR amplification and interpretation of results, specific to donkey DNA detection.

The ISO 20224 series consists of technical specifications that describe specific applications. New species DNA detection methods established in the future will be added as independent parts.

1 Scope

This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of donkey-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 donkey material derived from donkey (Equus asinus), mule (Equus caballus ♀ × Equus asinus ♂) and hinny (Equus caballus ♂ × Equus asinus ♀). The assay also detects the species zebra (Equus burchellii).

The target sequence is a partial fragment of the Equus asinus isolate Maral har breed Guanzhong donkey unplaced genomic scaffold, ASM130575v1 scaffold786, whole genome shotgun sequence (i.e. GenBank accession number NW_014638576.1)^[1], 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 (LOD_{95 %}).

2 Normative references

The following documents are referred to in the text in such a way that some or all of their content constitutes requirements of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.

3 Terms and definitions

For the purposes of this document, the terms and definitions given in ISO 16577 apply.

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Bibliography