ISO 20224-6 is the sixth part of a series of documents on molecular biomarker analysis. ISO 20224-6 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 the 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 %).
ISO 20224-6 on molecular biomarker analysis is relevant to:
A real-time polymerase chain reaction is a method to amplify DNA and make millions to billions of copies of a specific DNA sample from a very small sample of DNA.
ISO 20224-6 provides information on apparatus, procedure, preparation of the test portion/sample, preparation of DNA extracts, PCR setup required to perform PCR method for the qualitative detection of horse-specific DNA derived from food and feed.
As a user of ISO 20224-6, you can identify meat animal species from nucleic acid present in the ingredients of food and feed by polymerase chain reaction (real-time PCR) analytical method. ISO 20224-6 guides the user for PCR amplification and interpretation of results, specific to horse DNA detection. Overall ISO 20224-6 provides a qualitative and quantitative method for the detection of animal-derived biological materials in food and feed.
ISO/TS 20224-6