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The knowledge of Red blood cells polymorphism and blood group antigens prevalence at the local and regional levels is necessary for safe blood transfusion services. This study was aimed to estimate the prevalence of significant blood group phenotypes like ABO, Rh, and Kell among the Najran people of Saudi Arabia. The transfusion transmittable infection (TTI) rate and blood abnormalities among various blood types were assessed to ensure safe blood transfusion. ABO and Rh blood prevalence (n=970) and Rh phenotype polymorphism were estimated in over 531 unrelated donors. The blood samples were screened for certain TTIs like AHBC- Anti-hepatitis B core, HTLV-1- human T-lymphotropic virus type I, HCV- hepatitis C virus, HBsAg- Hepatitis B antigen, HIV- Human immunodeficiency virus, SIC- Sickle cell, MP- Malaria parasite, and SYP- Syphilis. The selected samples were also observed for blood abnormalities by performing a complete blood count (CBC). Out of 970 subjects, 966 were males, and only 4 were females. The O>A>B>AB blood groups were identified with 46.89, 29.3, 9.1, and 2.38% prevalence among Rh-positive phenotype. While in the Rh system, 87.6% and 12.3% of Rh positive and Rh negative was observed. Among 953 samples, the prevalence of seropositive donors was approximately 5.66%. The screening showed about 5.036, 0.104, 0.314, 0.209, 2.18, 0.104, and 0.209% positivity for AHBC, HCV, HBsAg, HIV, SIC, MP, and SYP respectively. Results found that the frequency of D, C, E, c, and e were 99.9%, 67.98, 25.8, 77.9, 98.49%, respectively, in over 531 subjects. The e allele was more prevalent in Najran city. After observing the variations in the CBC parameters among the donors, it was perceived that about 28.78, 99.9, 29.41, and 31.6% of blood abnormalities were noticed for O, AB, B, and A blood groups, respectively. For blood banks and transfusion services, which play a significant role in the medical care of the patient, awareness of the distribution of the blood group is essential. Increasing consistency of blood donation programs would improve both donor satisfaction and motivation for potential donations of blood in near future.