ISO TS 20224-2
Molecular biomarker analysis — Detection of animal-derived materials in foodstuffs and feedstuffs by real-time PCR — Part 2: Ovine DNA detection method
| Organization: | ISO |
| Publication Date: | 1 July 2020 |
| Status: | active |
| Page Count: | 24 |
| ICS Code (General methods of tests and analysis for food products): | 67.050 |
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).
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 %).
[1] BIGNON C., BINART N., ORMANDY C., SCHULER L.A., KELLY P.A., DJIANE J. Long and short forms of the ovine prolactin receptor: cDNA cloning and genomic analysis reveal that the two forms arise by different alternative splicing mechanisms in ruminants and in rodents. J. Mol. Endocrinol. 1997, 19(2), pp. 109-120
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