The total proteomes of Anisakis simplex s.s., A. pegreffii and their hybrid genotype have been compared by quantitative proteomics (iTRAQ approach), which considers the level of expressed proteins. A total of 1,976 proteins have been identified using public databases. One hundred ninety six proteins were found significantly differentially expressed; results of pairwise Log2 ratio comparisons among them were statistically treated and supported in order to convert them into discrete character states. This comparison selected thirty six proteins as discriminant biomarkers among A. simplex, A. pegreffii and their hybrid genotype; eighteen of these biomarkers, encoded by nine loci, are specific allergens of Anisakis (Ani s7, Ani s8, Ani s12 and Ani s14) and other (Ancylostoma secreted) is a common nematodes venom allergen.