【Objective】 To retrospectively investigate the antibody distribution and pregnancy outcome of pregnant women with Rh alloantibody in Guangzhou, and summarize the prevalence, diagnosis and treatment experience of hemolytic disease of newborn (HDN) caused by Rh alloantibody, so as to provide data for the prevention, diagnosis and treatment of Rh-HDN. 【Methods】 A total of 17 345 pregnant women in Guangzhou from January 2014 to December 2020 were selected for irregular antibody screening test.Those with Rh antibody were followed up for delivery, and the clinical and laboratory examination results of pregnant women and newborns were analyzed. 【Results】 A total of 71 cases (0.41%, 17/17 345) with Rh alloantibodies were detected.Among them, anti-D, anti-E, anti-Ec, anti-C, anti-Ce, anti-c, anti-e accounted for 26.76% (19/71), 46.48% (33/71), 9.86% (7/71), 7.04% (5/71), 5.63% (4/71), 2.82% (2/71) and 1.41% (1/71), respectively.Among the 71 pregnant women, 34 gave birth to children with HDN, with the total prevalence rate of 47.89%, among whom 100%, 78.94% and 42.42% were anti-c, anti-D and anti-E, respectively.And 71.43% (5/7) of the children who underwent transfusion were with anti-D.Although the yield rate of anti-E was the highest, it involved low morbidity and mild symptoms, which preferred to occur in the first fetus.No significant difference in gestational age, birth weight and the occurrence time of jaundice was notice between the anti-D group and anti-E group, but the total bilirubin of the anti-E group was lower while the Hb level were higher than those of the anti-D group (P<0.05). Two children died, and others were cured by phototherapy, albumin, IVIG and blood transfusion. 【Conclusion】 The publicity of Rh-HDN for early prevention and treatment should be strengthened to improve the cure rate and the prognosis.