Purpose: Meeting these requirements, a new fast allergen ELISA assay was developed and validated for 8 different allergens, namely: Almond, Casein, Cashew, Egg, Hazelnut, Macadamia nut, Mustard and Peanut.
Methods: All eight allergen ELISA test kits incorporate an extremely fast extraction procedure – using only extraction capsules containing a proprietary powdered buffer and hot water – of only 1 minute and short incubation times in the ELISA assay of only 10 minutes. Besides a low cross reactivity, the kits also showed a good performance in intra and inter-assay precision validation with variation being below 15% in almost every kit.
Results: The limit of detection (LOD) was calculated based on the mean blank value of 19 blank extractions plus three-fold standard deviation. All kits have LODs ranging in the low mg/kg level (Almond: 0.5 mg/kg; Casein: 0.2 mg/kg; Cashew: 1 mg/kg; Egg: 0.5 mg/kg; Hazelnut: 1 mg/kg; Macadamia nut: 1 mg/kg; Mustard: 0.5 mg/kg; Peanut: 0.5 mg/kg). Recoveries were ranging from 64 – 130% when kits were challenged to recover spiked allergens in difficult food matrices such as chocolate, milk drinks, cookies and ice cream. The kits also showed an equivalent performance in recovery when being compared to established ELISA assays from the market.
Significance: The validation showed that these new developed allergen ELISA assays are not only capable providing results in an extreme fast way, but also yield accurate results that you can rely on, making them suitable tools for the detection of allergens in many different foodstuff.