Molecular biomarker analysis — Detection of plant-derived materials in food by real-time polymerase chain reaction (PCR) — Part 2: Hazelnut DNA detection method

This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of hazelnut - specific DNA derived from food. PCR amplification reactions in which sufficient amounts of DNA are extracted from the relevant substrate can be used to detect the genetic material of hazelnuts, including hazelnut (Corylus heterophylla) and hazelnut (Corylus avellana). The assay also detects the Corylus heterophylla × Corylus avellana.
The target sequence is a partially fragment of the hazelnut (Corylus heterophylla) major allergen Cor h 1 gene and hazelnut (Corylus avellana) major allergen Cor a 1 gene (GenBank accession number EU195058.1 for Cor h 1 and GenBank accession number AF323975.1 for Cor a 1). The provided PCR assay for this target has an absolute detection limit of 5 copies per reaction, with ≥ 95 % reproducibility at this concentration (LOD95%).