*2.4. In Vitro Tests*

Total IgE and specific IgE Assay (ImmunoCAP 250; Phadia, Sweden) were performed in patients with positive SPT to evaluate the major inhalant allergen. The level of awareness towards the main inhalant allergen was increased using the CRD. We evaluated IgE antibodies to Phl p1 (Timothy grass), Phl p5 (Timothy grass), Bet v1 and Bet v2 (Betula verrucose), Amb a1 (Ambrosia), Art v1 (Mugwort), Par j2 (Wall pellitory), Ole e1 (Olea europea), Cup a1 (Cupressus arizonica), Fel d1 (cat), Can f1 (dog), Der p1 (House dust mite), Der p2 (House dust mite), and Alt a1 (Alternaria alternata). IgE levels were considered positive at the level ≥0.35 kUA/l. Patients with IgE antibodies to Bet v2 were excluded to rule out profilin allergy [31–33].
