This study aimed to characterize anti-A and anti-B hemolysins, IgM, and IgG titers in Thai blood donors. Altogether, 300 serum samples from group O donors at the National Blood Centre, Thai Red Cross Society, were screened for anti-A and anti-B hemolysins and treated with 0.01 M dithiothreitol to characterize IgM and IgG titers by standard tube technique. Antibody titers were compared with hemolysis grade. Male and female ratio = 1:1.3 and ages ranged from 17 to 60 years. The overall prevalence of anti-A and anti-B hemolysins was 69%. Anti-A and anti-B hemolysins comprised 18.3% and 16.7%, respectively and 34% had both antibodies. High titers of anti-A hemolysins were associated with females (P< 0.05), and only anti-B IgM titers were associated with age (P< 0.05). Interestingly, the association of anti-A IgM titers, anti-A IgG titers, and hemolysin grade was demonstrated (P< 0.05). A significant association between hemolysin grade and anti-B IgM titers was found (P< 0.05). The prevalence of anti-A and anti-B hemolysins and high titers of IgM and IgG in Thais are high. Hemolysin grade showed significant associations with IgM titers; therefore, when providing ABO-incompatible platelet transfusion, especially for female plateletpheresis donors, IgM high titers of anti-A and anti-B screening is suggested.