Prehrambeni proizvodi — Detekcija alergena u hrani molekularno-biološkim metodama — Deo 8: Kikiriki (Arachis hypogaea), lešnik (Corylus spp.), orah (Juglans regia) i indijski orah (Anacardium occidentale) — Kvalitativna detekcija specifičnih DNK sekvenci u hrani pomoću PCR u realnom vremenu
Foodstuffs - Detection of food allergens by molecular biological methods - Part 8: Peanut (Arachis hypogaea), hazelnut (Corylus spp.), walnut (Juglans regia) and cashew (Anacardium occidentale) - Qualitative detection and quantitative determination of specific DNA sequences in food by real-time PCR
This document specifies a method for the qualitative detection and quantitative determination of the species-specific DNA of peanut (Arachis hypogaea), hazelnut (Corylus spp.), walnut (Juglans regia) and cashew (Anacardium occidentale) in food of animal and plant origin, using real-time PCR, in the context of allergen analyses.
The method was previously validated in an interlaboratory study (ring trial) and applied to DNA extracted from samples that consist of defined proportions of peanut, hazelnut, walnut and cashew in rice biscuits, cooked sausage, sauce powder, vegan cookie and veggie burger (powder).
The limit of detection (LOD) of each real-time PCR system has been determined experimentally to be about 1 mg nuts per kg food. The limit of quantification (LOQ) of the method was tested using different food samples spiked with peanut, hazelnut, walnut and cashew (see 8.5). The ring trial data showed that 5 mg of nuts per kilogram of food can be reliably determined. A good quantification is possible, especially if the source of the allergenic ingredient (e.g. roasted peanuts) is known and can be used for the calibration of the method.
