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Many tree nuts are considered to be a serious problem in food safety, because of the presence of causative factors in IgE-mediated food allergies. Among these, hazelnut (Corylus avellana L.) seeds are largely used in a range of confectionery products and contain many well-characterized allergens. DNA-based methods and ELISA tests may prove to be useful to assess the presence of hidden ingredients in foods. The aim of this work was the development and validation of a species-specific SYBR Green I real-time PCR protocol for the detection of hazelnut in foods. A novel efficient primer pair on the Cor a 8 genomic coding region was designed by preparing a plasmid vector-based internal reference standard to calibrate the PCR. A good sensitivity, down to 20 (genomic) and 15 (plasmid) DNA copies, was established. All of the commercial samples considered in our study (containing hazelnut as ingredient or as a potential trace cross-contamination) were effectively amplified by PCR, showing a perfect correspondence with an ELISA commercial test, employed as a reference standard method.

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Summary Cor a 1, Cor a 8 and Cor a 11 are three important allergens in hazelnut. Aim of this work was to set up a method to quantitatively measure differences in their transcription levels. For the expression study, three different models were set up, taking also into account the ripening stage and the year of harvest. A gene transcription analysis by relative quantification through Real‐Time PCR has been developed to measure the transcription profile of hazelnut allergens. Differences in the gene transcription levels of Cor a 8 and Cor a 11 were found, both when comparing different cultivars among themselves and when comparing different years of harvest and ripening stages in the “Tonda Gentile Trilobata” (TGT) cultivar. Major allergen Cor a 1 gene did not display any statistically significant variation in its transcript abundance. Principal Components Analysis (PCA) was finally performed to better elucidate the classification of samples.