Lupine flour, protein, and fiber have become common ingredients in food products. The association of lupine-related allergic incidents with peanut allergy is a cause for concern as the latter may bring about severe reactions. In this study, a hybridization probe-based real-time PCR assay for the detection of lupine DNA in foods was developed. Particular attention was paid to the specificity of the method, which was verified by analysis of DNA extracts from more than 50 potential food ingredients such as legumes, cereals, seeds, nuts, spices, fruits, and meat. The limit of detection of the method was determined as 0.1 mg/kg. The successful detection of the presence/absence of lupine DNA in 20 samples proved the suitability of the assay for the analysis of frequently encountered food matrices.
The simultaneous detection of DNA from different allergenic food ingredients by a ligation-dependent probe amplification (LPA) system is described. The approach allows detection of several targets in a one-tube assay. Synthetic oligonucleotides were designed to detect DNA from peanuts, cashews, pecans, pistachios, hazelnuts, sesame seeds, macadamia nuts, almonds, walnuts and brazil nuts. The specificity of the system was tested with DNA from more than 50 plant and animal species. The sensitivity of the method was suitable to detect allergenic ingredients in the low mg kg(-1) range. The limit of detection (LOD) for single allergens in different food matrices was 5 mg kg(-1). The novel analytical strategy represents a useful tool for the surveillance of established legislation on food allergens within the European Union.
Appropriate detection methods have to be provided to assure the compliance with the recently established regulatory provisions concerning the labeling of allergens in food. Therefore, a novel real-time polymerase chain reaction (PCR) system for the specific and sensitive detection of cashew nut (Anacardium occidentale) was developed. Specificity was checked against DNA from 56 plant and animal species to avoid cross-reactivity to phylogenetically related and other food-relevant organisms. The absolute limit of detection (LOD) was determined to be 0.5 pg genomic cashew DNA and 10 copies, respectively, and the practical LOD examined exemplarily for pesto Genovese was 2 mg/kg. In addition, analysis of different retail samples was performed to demonstrate the suitability of the new assay for manifold applications.
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