Molecular biomarker analysis. Detection of animal-derived materials in foodstuffs and feedstuffs by real-time PCR. Horse DNA detection method
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
horse-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 horse
material derived from domestic horse (Equus caballus), mule (Equus caballus ? × Equus asinus ?), hinny (Equus caballus ? × Equus asinus ?) and zebroid (Equus caballus × Equus simplicidens). The assay also detects the species
Przewalski’s horse (Equus przewalskii) and zebra (Equus burchellii).
The target sequence is an Equus caballus isolate Twilight breed thoroughbred
chromosome 28, EquCab3.0, whole genome shotgun sequence (i.e.
GenBank accession number NC_009171.3)[ 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 %).
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