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Background and Objectives Alloimmunization with the prevalence rate of 60% is a common problem in chronically transfused patients. This problem has been reported to be less common in transfused hospital-based patients, amounting to 1-2 percent. Alloimmunization can lead to some difficulty varying from delay in the provision of similar blood types to delayed hemolytic transfusion reactions. The aim of this study was to analyze alloimmunization against RBCs among non-chronically transfused patients in Namazi Hospital in Shiraz. Materials and Methods Retrospective analysis of 3487 non-chronically transfused patients was conducted with antibody screening test and identification of patients' alloantibodies with the panel cell test. For patients with alloantibody, the data for sex, age, and history of transfusion and surgery were collected. Finally, the data were analyzed by SPSS software v. 15 using chi-square (CI= 95%). Results Twenty-eight out of 3487 patients had alloantibody (prevalence rate of 0.8%). The most common clinically significant alloantibodies found were anti-K (23%), anti- E (15%) and anti-C (11%). The most common clinically significant alloantibodies identified in males and females were anti-K and anti-C, respectively. Conclusions Alloantibody prevalence rate did not show any correlation with age and sex and was more common in patients with surgery history and transfusion record. Important factors contributing to the higher prevalence of the above alloantibodies are the higher prevalence of the related antigens in the population, higher antigenicity power, and the lack of crossmatch and grouping for the antigens before blood transfusion. [ABSTRACT FROM AUTHOR]