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.