【Objective】 To retrospectively analyze the irregular antibodies in 6 blood group systems other than the Rh blood group system in 53 pregnant women and analyze its correlation with the occurrence of hemolytic disease of the newborn(HDN). 【Methods】 19 473 pregnant women were screened for irregular antibodies by microgel detection technology combined with anti-human globulin (IgG+ C3d), and the positive samples screened out were further confirmed to understand the types and titers of irregular antibodies. Irregular antibody type determination experiment: IgG type irregular antibody titer was determined after mercaptoethanol (2-Me) inactivated the serum of the irregular antibody positive specimen, and then IgG and IgM type were determined by comparing the titer levels of irregular antibody. Three hemolysis tests and total bilirubin tests were performed on umbilical cord blood during delivery to analyze the level of jaundice and the occurrence of HDN. 【Results】 53 cases of irregular antibodies other than the Rh blood group system were detected in 19 473 pregnant women, with a positive rate of 0.27%, mainly MNS and Lewis blood group system.The incidence of HDN was 39.6% (21/53). There were 27 cases of IgM, 7 IgG, and 19 IgM + IgG. Comparison of total bilirubin detection between the low titer group (≤8) and the high titer group (>8) : the latter was significantly higher than the former (P<0.05); IgG antibody subtypes: IgG1 of the latter significantly increased (P<0.05), and so was IgG3 in former (P<0.05). There was a significant positive correlation between IgG1, IgG3 and total bilirubin. The area under the curve of IgG1+ IgG3 for HDN diagnosis, the sensitivity and specificity were 0.953, 0.900, and 0.967, respectively. 【Conclusion】 Other than Rh blood group system, irregular antibodies are mainly distributed in MNS and Lewis blood group system. The incidence of HDN is higher in Kell, Duffy and Kidd blood group systems after producing irregular antibodies. Non-antibody types are mostly IgM type or IgM + IgG mixed, and the incidence of HDN is not high; Patients with poor maternal history, either high or low titer, can be classified into IgG1 and IgG3 in early stages, and those with Abnormal results should be included into the perinatal management of high-risk women with regular checking.