Background: Malaria is a disease transmitted by vectors and caused by unicellular Plasmodium parasites. Malaria pathogenesis is associated with the ABO phenotype. However, there is little information on the frequency of malaria disease and its relationship with the ABO blood group in the study area. Therefore, the purpose of this study was to determine the prevalence of malaria infection and its association with the ABO blood group at Woldia Comprehensive Specialized Hospital.Method: An institutional‐based cross‐sectional study was conducted from December 3, 2022, to February 30, 2023. Convenient sampling was used for selecting the study participants. To identify malaria parasites, thick and thin blood films were made. Additionally, blood was drawn to identify the ABO blood group type. Before being analyzed with SPSS software Version 27, the data was coded and entered into EpiData Version 3.1. To ascertain the variable’s association, a logistic regression was done.Results: Out of 192 patients that attended Woldia Comprehensive Specialized Hospital, 16 (8.3%) were found to be infected with Plasmodium parasites using microscopy. Among them, 9 (4.7%), 5 (2.6%), and 2 (1.0%) had Plasmodium falciparum, Plasmodium vivax, or mixed infections, respectively. As a result, 30.7%, 25.5%, 24.5%, and 19.3% of the participants had blood types A, B, AB, and O, respectively (AOR = 2.359, 95% CI: 1.03–12.289, p = 0.03).Conclusion and Recommendation: The total number of microscopically confirmed malaria parasites was 8.3%. P. falciparum was dominant over P. vivax. Individuals with blood group O were less likely to get severe malaria than those with other blood groups. Based on the findings of this study, we recommend that additional studies investigate the probable relationship between the ABO blood group phenotype and malaria infection.