BackgroundKnowledge of the ABO and RhD group distribution is essential for blood banks inventory and assuring quality blood transfusion services. The objective of this study was to determine the frequency of ABO and RhD phenotype, allele, and genotype among blood donors at North Gondar District Blood Bank from 2010 to 2012, Northwest Ethiopia.MethodsThe data of the current study were obtained from registration logbooks of blood donors registered. The ABO and RhD grouping was done by using commercially available monoclonal antibodies (anti-A, anti-B and anti-D) by slide methods. Results with no agglutination by anti-D antibody were confirmed using anti-human globulin test. Descriptive statistics were analyzed using SPSS version 20. The allele and genotype frequency of the donors was determined by Hardy–Weinberg equilibrium assumption. The difference between the observed and expected frequency was tested by online Chi-square calculator. P-value of <0.05 was considered statistically significant.ResultsAmong 6471 blood donors, 82.1%, 94.1% and 55.4% were males, replacement donors and in the age group of 21–30 years, respectively. Blood group O (47.04%) and blood group AB (4.81%) were the dominant and least common, respectively. The distribution of the RhD negative blood group was 5.76%. The distribution of A, B and O alleles was 0.1714, 0.1433 and 0.6859, respectively. Moreover, the genotype frequency of AA, AO, BB, BO, AB and OO was 0.0294, 0.2350, 0.0205, 0.1966, 0.0491 and 0.4704, respectively. The genotype frequency of DD, Dd and dd was 0.5774, 0.3649 and 0.0576, respectively. The result showed that there was no statistically significant difference between observed and expected allele and genotype frequency (P-value >0.05).ConclusionBlood group O and AB were the most and least prevalent, respectively. The allele and genotype frequency of the population was fulfilled the Hardy–Weinberg equilibrium assumption. This finding might be useful for blood transfusion services.