BACKGROUND New in vitro diagnostic methods for IgE-mediated drug allergic reactions, such as basophil activation test and antigen specific sulfidoleukotriene test, have proven their usefulness in patients with positive skin tests. OBJECTIVE To assess the usefulness of basophil activation test and antigen specific sulfidoleukotriene test in the diagnosis of patients with IgE-mediated allergy to Betalactam antibiotics and negative skin tests. METHODS The 23 patients included in the study underwent basophil activation test, antigen specific sulfidoleukotriene test and specific IgE. The patients were classified into three groups. patients with positive specific IgE. patients with a unique immediate reaction to Betalactams, negative specific IgE and positive oral provocation tests. And Group C: patients with at least two immediate reactions induced by Betalactams and negative specific IgE. RESULTS The sensitivity/specificity of the different tests are: basophil activation test 39.1 %/93.3%, antigen specific sulfidoleukotriene test 22.7%/83.3%, specific IgE 21.7%/86.7%. The joint use of the three tests allows diagnosis of 60.9% of the patients. CONCLUSIONS In vitro diagnostic tests, especially basophil activation test, are very important tools in the diagnosis of patients with IgE-mediated allergy to Betalactams and negative skin tests, avoiding performance of potentially dangerous oral provocation tests in a high percentage of cases.