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Hidden allergens in food: Peanut detection by an official food safety laboratory

Clara Tramuta, Daniela Manila Bianchi, Elisa Barcucci, Sandra Fragassi, Samantha Lupi, Enrico Arletti, Melissa Bizzarri and Lucia Decastelli

Food safety laboratories rely on validated methods that detect hidden allergens in food to ensure the safety and health of allergic consumers. Here we present the test results for the validation and accreditation of a real time PCR assay (RT-PCR: SPECIAL finder MC Peanut, Generon), for the detection of peanut traces in food. The method was tested on five classes of food matrices: bakery and pastry products, meats, ready-to-eat and dairy products, grains and milling products. Blank samples were spiked starting with the peanut samples (Arachis hypogaea), lyophilized and stored at -18 °C, at a concentration of 1000 ppm. Serial dilutions were then prepared with the DNA extracted from the blank samples to final concentration of 0.5 ppm. Results: LOD 0.5 ppm in grains and milling products, ready-to-eat, meats, bakery and pastry products (range Ct 27-34); LOD 2.5 ppm in dairy products (range Ct 25-34). To determine the exclusivity parameter of the method, the ragù matrix was contaminated with Prunus dulcis (almonds), Glycine max (soy), Sinapis alba (mustard), Apium graveolens (celery), Allium cepa (onion), Pisum sativum (peas), Daucus carota (carrots), and Theobroma cacao (cocoa). No cross-reactions were observed. Method accuracy was rated satisfactory for sensitivity (96%) and specificity (100%) and was validated. It was found adequate for the needs of the laboratory as it meets the purpose for which it was applied.

This study was funded in part within a project of the Italian Ministry of Health (IZS PLV 02/19 RC).