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Background and Objectives: The occurrence of hemolysis due to transfusion of ABO plasma-incompatible platelets (PLTs) is challenging. There has been no consensus for critical antibody titers in the transfusion community. This study was conducted to understand the trends of anti-A and anti-B antibody titer levels in O group donors and to identify any specific patterns of distribution in relation to age and gender. Materials and Methods: A total of 1635 Group O PLT donors were randomly selected for this prospective study. Serial 2-fold doubling dilutions were prepared for each sample to calculate the titer of anti-A and anti-B in a standard 96 well micro-plate. Tube technique was used for comparison with the microplate method for 100 samples. Results: Out of 1635 donors, 1430 (87.46%) were males and 205 (12.54%) were females. The median titer for anti-A and anti-B was 128 with range from 4 to 2048. Spearman's correlation coefficient for microplate versus tube technique was estimated to be 0.803 (P < 0.01, two-tailed). 57.12% and 51.19% of all donors had titers ≥128 for anti-A and anti-B, respectively. The geometric mean of anti-A and anti-B was 155.7 and 137.28, respectively. The titers were significantly higher (P < 0.001) in female donors. An inverse relation between titer levels and age was seen. Conclusion: Microplate can be used to perform titers in resource-constrained settings. Screening for critical titers in O group donors is essential as they are more implicated in hemolytic transfusion reactions. In the absence of a global consensus on this topic, institutes may need to formulate their own guidelines on handling ABO plasma-incompatible PLT transfusions. [ABSTRACT FROM AUTHOR]