The rising popularity of undercooked or raw seafood containing larvae of theAnisakisparasite has led to issues of public health concern due to allergic manifestations. We conducted an observational study on the use of an innovativeAnisakisallergy diagnostic algorithm in a convenience sample of 53 allergic outpatients recruited in Western Sicily, between April 2021 and March 2022. We included individuals with an anamnesis suggestive of IgE sensitization toAnisakisreporting clinical manifestation in the last month due to allergic reactions after eating fresh fish, or in subjects at high exposure risk with sea products while abstaining from fish ingestion, excluding those with documented fish sensitization. Outpatients were tested via Skin Prick Test, IgE-specific dosage and Basophil Activation Test (BAT). Twenty-six outpatients were diagnosed withAnisakis, while 27 with Chronic Urticaria (CU). We found a seven-fold excess risk forAnisakis(p4) positivity in theAnisakisallergic outpatients, as compared to the CU ones. BAT showed the best diagnostic accuracy (92.45%) and specificity (100%), while specific IgE toAscaris(p1) documented the best sensitivity (92.31%) but a very low specificity (37.04%). In conclusion, our findings may represent a potentially useful contribution to the future development of updated clinical guidelines.