Background: Blood transfusions are a life-saving medical procedure, yet they carry the risk of Transfusion-Transmitted infections, including Transfusion-Transmitted Malaria. This risk is heightened in Sub-Saharan Africa and Ethiopia due to a large pool of asymptomatic, low-parasitemia donors who evade conventional screening. The risk is compounded by the absence of routine malaria screening in Ethiopian blood banks and the limited sensitivity of available diagnostic methods. Examining the link between malaria and host genetic factors, such as ABO blood group, could support the development of targeted, resource-appropriate screening strategies. Objective: To determine the prevalence of malaria among blood donors and its association with ABO blood group variants using different diagnostic methods in Bahir Dar, Northwest Ethiopia. Methods: A facility-based cross-sectional study was conducted among 423 blood donors at five mobile blood collection sites in Bahir Dar, Northwest Ethiopia,from January–July 2026, using systematic random sampling. Three milliliters of Blood samples were used for screeningPlasmodium infection using BIOCREDIT pfLDH/pvLDH RDTand microscopy for 423 samples, and PCR (nested PCR and qPCR) on a sub-cohort of 221 samples. Sociodemographic and relateddata were collected using a structured questionnaire and analyzed using SPSS version 25 and RStudio software with descriptive statistics, Cohen's kappa, chi-square, and logistic regression (p<0.05 significant) Results: The malaria prevalence was 3.5% (15/423) by microscopy, 4% (17/423) by RDT, 4.5% (19/423) by composite RDT and microscopy, and 4.5% (9/202)by nested and qPCR.RDT and microscopy showed sensitivity of 56% and 52% respectively with a tie overall accuracy of 93.7%.P. vivaxwas predominant, with zero prevalence of P. malariae andP. ovale. Blood group O was most prevalent (47%; 199/423) and showed significantly higher malaria positivity (10.1%) and odds of infection (AOR=3.08, 95% CI: 1.14–9.11). Rural residence, malaria history, and travel to endemic areas independently increased infection odds, while Rh-positive status and ITN use were protective. Conclusion and Recommendation: This study revealed a considerable burden of asymptomaticPlasmodium infection among blood donors in Bahir Dar.Despite WHO recommendations, malaria screening is not currently integrated into routine blood donation practices. Therefore, incorporating conventional screening methods and more sensitive molecular tools is essential to enhance transfusion safety.