ISO/NP 20224-13.2

Molecular biomarker analysis — Detection of animal-derived materials in foodstuffs and feedstuffs by real-time PCR — Part 13: Rainbow trout DNA detection method ISO/NP 20224-13.2

Scope

This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of rainbow trout (Oncorhynchus mykiss)-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 rainbow trout material. The assay also detects the gila Trout (Oncorhynchus gilae) and westslope cutthroat trout (Oncorhynchus clarkii lewisi) which are endangered fish species.
The target sequence is a partial fragment of rainbow trout ATPase 6 gene (i.e. GenBank accession number KP013084.1)[1],which is part region of the rainbow trout mitochondrion genome. The provided PCR assay for this target has an absolute limit of detection of 5 copies per reaction, with ≥ 95 % confidence at this concentration (LOD95 %).