Collaborative trial validation of a new multiplex real-time PCR to sensitively detect allergenic nuts in food

Abstract In this article, we present a multiplex real-time PCR method for simultaneous, sensitive and specific detection semi-quantitative estimation of the allergenic species peanut, hazelnut, walnut cashew in food. Due to use multicopy target sequences, very ingredients was possible. The validated in-house as well by collaborative trial with 12 laboratories. Within ring trial, 0.64 mg/kg (i.e. approx. 0.1–0.2 mg peanut tree nut-derived protein/kg) could still be detected processed cookie matrix, confirmed results incurred, samples spiked at low levels between 0.9 50 corresponding ingredient (peanut, nut). addition, revealed good precision data. With regard quantitative analysis though, insufficient recovery data (bias) were determined some cases, resulting measurement uncertainties more than 50%.