ISO/NP TS 20224-2.1

Molecular biomarker analysis — Detection of animal-derived materials in foodstuffs and feedstuffs by real-time PCR — Part 2.1: Ovine DNA detection method

Scope

This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of ovine-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 ovine material derived from sheep (Ovis aries). Cross detection of the wild animals Aoudad (Ammotragus lervia), Bighorn sheep (Ovis canadensis), Blue sheep (Pseudois nayaur), Hartebeest (Alcelaphus buselaphus), Hirola (Beatragus hunteri), Nilgiri tahr (Nilgiritragus hylocrius), Northern chamois (Rupicapra rupicapra), Siberian ibex (Capra sibirica) and Takin (Budorcas taxicolor) is observed.
The target sequence is a partial fragment of the ovine nuclear prolactin receptor short form mRNA gene (PRLR) (i.e. GenBank accession number AF041979.1)[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 (LOD95 %).