Data Analysis
Statistical analyses were performed by GraphPad Prism 8. Comparison of general characteristics of sensitized and non-sensitized subjects was evaluated by chi-square test, t test or Mann-Whitney U test where appropriate. Differences in sIgE among different fishes and components were compared using Friedman’s ANOVA post-hoc test. Correlation analysis was performed with Pearson’s correlation test. ED10 values for salmon and grass carp (doses at which an allergic reaction would be elicited in 10% of the population in oral food challenge) were calculated as described 24 . The survival package 29 fin R30 was used for interval-censoring survival analysis. 95%CIs for the ED10 values were calculated by taking the 2.5th and 97.5th percentiles of the bootstrap distribution of ED10 values generated using 10,000 bootstrap samples. IgE-binding epitopes were defined for peptides with average spot signals (1) ≥1000 a.u., (2) > 3-fold of the signal from anti-human IgE secondary antibody, (3) > 3-fold of the signal from control serum sample, and (4) significantly higher signal intensity of allergic samples than tolerant samples (i.e. tolerated salmon or tolerated all fishes in food challenge) based on 1% false discovery rate (FDR) in Mann-Whitney test. Protein sequence alignment was performed on Clustal Omega 25 . A p-value of <.05 was considered statistically significant.