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Canine atopic dermatitis (CAD) is a chronic, inflammatory, and pruritic disease of the skin resulting from the loss of the epidermal barrier, sensitization, and exacerbated production of IgE antibodies mainly directed against environmental allergens, especially to house dust mites. To select specific allergen immunotherapies with high efficacy, there are necessary studies with house dust mite allergens to improve both serological and intradermal tests. Therefore, the objective of this study was to evaluate the seroprevalence of IgE against Der f 2, Zen 1, and crude Dermatophagoides farinae allergens in dogs with AD in the State of São Paulo, Brazil. The sera of 85 dogs with clinically confirmed atopic dermatitis from the State of São Paulo (Brazil) was collected. In addition, an indirect ELISA test was conducted to detect allergen-specific serum IgE. IgE seropositivity was observed in 97.5% of the dogs for Der f 2, 95.0% for Zen 1, and 92.5% for the crude mite allergens. Due to this high prevalence of IgE specific to these allergens, we suggest that Der f 2 and Zen 1 can be considered major allergens for dogs in the State of São Paulo., Competing Interests: Declaration of Competing Interest We declare that Dr. Toshihiro Tsukui and Ms. Miyuki Kageyama are employees of Zenoaq., (Copyright © 2023 Elsevier Ltd. All rights reserved.)