Objective To investigate the positive rate and distribution of unexpected antibody in non-remunerated voluntary blood donors. Methods An accidental antibody screening on 2 100 qualified component blood samples collected by Xuancheng People's Hospital from October 2022 to December 2023 before clinical infusion was performed,and then the red blood cell specific antibodies in blood donors with positive screening results was identified. Results A total of 2 100 non-remunerated voluntary blood donors were screened,including 1 271 males,accounting for 60.52%,and 829 females,accounting for 39.48%.A total of 12 cases of accidental antibodies were detected,with a positive rate of 0.57%.The positive rate of accidental antibodies in males was 0.16%,which was lower than that in females (1.21%) (χ_corrected²=7.958,P=0.005).The positive rate of accidental antibodies in blood donors with a history of blood transfusion was 3.92%,which was higher than that in blood donors without a history of blood transfusion (0.49%) (P_Fisher=0.033 ).Among the 12 cases of accidental antibody positive,11 cases were specific antibodies,including anti-E (5 cases),anti-M (3 cases),anti-c (2 cases) and anti-Lea (1 case).One case of non-specific antibody was cold autoantibody. Conclusion It is of great significance to carry out unexpected antibody screening and identification of voluntary blood donors before blood transfusion,which can reduce cross-matching incompatibility,and the risk of blood transfusion,and ensure the safety of clinical blood use.