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Objective: Blood transfusion is a common lifesaving treatment but it is often complicated with alloimmunization. Previously studies in Greece have concentrated on alloimmunization in multiply transfused thalassemic patients or antenatal women. However, the relative frequency of red blood cell (RBC) alloantibodies in the general patient population has not been studied so far. The aim of the present retrospective study was to estimate the prevalence and specificity of RBC alloantibodies in a large cohort of patients in two general hospitals and their association with age, sex, and the patients' clinic of hospitalization. Materials and Methods: Data from 2012 to 2016 from the 'Sismanogleio' and 'Thriasio' general hospitals in Athens, Greece, were studied retrospectively. Statistical analysis was performed with SAS for Windows 9.4. Results: Six hundred twenty-six patients (626/53800, 1.16%) were alloimmunized for one or more alloantibodies. The mean age was 67.99+-17.56 years. Most antibodies were found in women [62.66% (438/699) in women vs. 37.34% (261/699) in men (p=0.0007)], while the vast majority of antibodies (66.81%) were found in patients aged 61-90. The most frequent antibody was anti-Kell (26.61%), followed by anti-E (16.02%), anti-D (15.02%), anti-Jka (5.87%), and anti-M (5.72%). Anti-C (81.48%, n=27) and anti-Cw (54.17%, n=24) tended to be found more often in patients with multiple antibodies. Most alloimmunized cases were found in general surgery (42.65%) and internal medicine departments (38.66%). Conclusion: According to our results, the alloimmunization data in a general patient population in Greece were consistent with the majority of studies in the international literature. Whether a strategy at national level needs to be directed towards extending matching for the whole population or towards applying sensitive and compulsory indirect antiglobulin tests before any transfusions in order to efficiently prevent alloimmunization remains an issue of debate.