Development and validation of a duplex real-time PCR method to simultaneously detect potentially allergenic sesame...

The paper describes the development and validation of a duplex real-time PCR method allowing the simultaneous detection of traces of potentially allergenic sesame and hazelnut in food. For the detection of sesame and hazelnut, the genes coding for two major allergenic proteins, Ses i 1 and Cor a 1, were selected. The duplex real-time PCR assay did not show any cross-reactivity with 25 common food ingredients from sesame and/or hazelnut containing foods. Analysis of serially diluted sesame/hazelnut DNA resulted in good linearity up to a dilution of 1:10000 (corresponding to 10 pg microL(-1) or 50 pg). Sesame and hazelnut could be detected in blank whole meal cookies which had been spiked with 0.005% sesame and 0.005% hazelnut. The applicability of the real-time PCR assay for determining sesame and hazelnut in different food matrices was investigated by analyzing 30 commercial foodstuffs comprising salty snacks, cookies, chocolates, creams, mueslis and muesli bars.