PD ISO/TS 20224-7:2020

This standard PD ISO/TS 20224-7:2020 Molecular biomarker analysis. Detection of animal-derived materials in foodstuffs and feedstuffs by real-time PCR is classified in these ICS categories:

• 67.050 General methods of tests and analysis for food products

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 %).