Background/Aim. Pregnancy-induced red blood cell (RBC) alloimmunization is important not only because of the possi-ble negative effects on subsequent pregnancy outcomes in case the fetus carries the antigen but also because of the optimal transfusion management in cases of obstetric hemorrhage. Timely detection of RBC antibodies is part of testing, preven-tion, and treatment strategy, aimed at achieving better out-comes for alloimmunized mothers with an affected fetus. The aim of the study was to determine the frequency and specifici-ty of alloantibodies among pregnant women from the South Bačka District, Serbia, with special attention to the incidence of anti-D alloantibodies. Methods. A retrospective study was conducted in the Blood Transfusion Institute of Vojvodina and covered the period from January 1, 2010, to December 31, 2019. Screening and antibody identification were performed by an indirect antiglobulin test in gel-microcards (ID-Card Liss/Coombs) with two test RBC (ID-DiaCell I-II screening cells, Bio-Rad, Cressier, Switzerland) on an automated system (IH-500, Bio-Rad). Results. Among 25,694 tested pregnant women, 1.38% were actively immunized, while 1.12% of women acquired antibodies in the current pregnancy. Among 3,622 (14.09%) RhD-negative women, 1.77% produced anti-D antibodies during the ongoing pregnancy. Distribution of antibody specificity was: anti-D 23.34%, anti-M 11.85%, anti-E 9.41%, anti-K 9.41%, anti-C 5.92%, anti-Fyb 5.92%, anti-c 3.13%, anti-S 3.13%, anti-Lea 3.13%, anti-Leb 3.13%, anti-Cw 1.75%, anti-Jka 1.40%, anti-P 1.05%, anti-Lub 0.70%, anti-Fya 0.35%, autoantibody of undetermined specificity 0.70%, and irregular antibodies of undetermined specificity 15.68%. Con-clusion. Immunoglobulin prophylaxis has led to a significant reduction in the frequency of D-alloimmunization among pregnant women in the South Bačka District over the last ten years. However, the incidence of anti-D antibodies is still significantly higher than in published data for developed countries. We also identified the other, less commonly pre-sent, clinically significant antibodies. There is a need to intro-duce uniform recommendations for immunohematological testing in pregnancy on the territory of the Republic of Serbia in accordance with modern requirements.