Objective: The distribution ratio and gene frequency of Rh blood group main antigen phenotype in some patients and donors in Nanjing were compared and analyzed to explore the feasibility of Rh blood group system main antigen coordination infusion and evaluate its effect, laying a foundation for accurate and safe blood transfusion. Methods: From January 2020 to May 2021, a total of 126,782 Rh blood group system D, C, c, E and e antigens were detected by microcolumn gel method in 93,910 patients in our hospital and 32,872 red blood cells from blood donors sent to our hospital. The phenotypic distribution and gene frequency of Rh antigen were calculated and the difference of detection rate was determined by Chi-square test. The distribution of Rh blood group was evaluated by Hardy-Weinberg equilibrium law. The blood transfusion information management system of our hospital recorded and stored the Rh blood group results of patients and donors respectively, and selected Rh homogenous or compatible donor blood bags for matching test before cross-matching. Results: The positive rate of RhD antigen in 93910 patients was 99.5% (93453/93910). A total of 15 phenotype were detected in 126782 Rh blood groups. The main phenotype rates for DCCee, DCcEe, DCcee, DccEE, DccEe were 42.27%(53596/126782), 35.70%(45267/126782), 9.01%(11423/126782), 7.56%(9587/126782), 3.81%(4834/126782),respectively, and 7 cases of rare phenotype DCCEE were detected. Rh phenotype distribution conforms to Hardy-Weinberg law. D antigen frequency was 99.57%, C antigen frequency was 88.03%, c antigen frequency was 57.06%, E antigen frequency was 48.00%, e antigen frequency was 92.18%. D gene frequency was 0.9348, C gene frequency was 0.6548, c gene frequency was 0.3452, E gene frequency was 0.2790, e gene frequency was 0.7209. The haploid DCe frequency was 0.6369, and the second is DcE with a frequency of 0.2724. There was no difference in Rh phenotype and gene frequency between patients and donors (P > 0.05). After 27889 Rh infusion times, the positive rate of antibody in Rh blood group system decreased from 48.61% (198/408) to 30.25% (108/357) (P < 0.05). Conclusion: There is no difference in Rh phenotype and gene frequency between patients and donors. It is feasible to perform routine Rh coordinated infusion based on ABO homotype in patients. Rh coordination infusion can significantly reduce the incidence of irregular antibody positive and improve the transfusion effect.