Abstract:To solve the shortage of hazelnut reference material for food allergen detection，selected hazelnut kernels were crushed，degreased by n-hexane，and sieved to make loosely degreased fine powder.Quantitative real-time polymerase chain reaction（qPCR）and enzyme-linked immunosorbent assay（ELISA）were used for confirmation，and the effects of degreasing on the characteristic values of hazelnuts before and after degreasing were compared.The uniformity，stability，and fixed value analyses were carried out.The qualified standard sample of defatted hazelnut powder was prepared and used to verify the detection limit of qPCR and ELISA.The results showed that the Ct value of Oleosin gene in the hazelnut powder sample before degreasing was 26.22±0.47，and that after defatting was 25.80±0.31.After analysis of variance（ANOVA），under the 95% confidence probability，the F ratios calculated by the two methods were 2.17 and 2.10，respectively，which were both less than the F-critical value F0.05(14，15)=2.42，showing a good uniformity.The|b1|of the measured values of the two methods were 0.011 67 and 0.000 24，respectively，which were less than t (0.95，n-2)×s（b1），and the statistical pvalues were 0.08 and 0.054（p>0.05），respectively.The p-values of the between-group statistics were 0.14 and 0.27（p>0.05），respectively，showing a good stability.The ELISA method had a better detection sensitivity than qPCR，detecting hazelnut residues at 0.000 1% in food.