Objective To explore the early diagnostic and treatment value of combined detection of direct antiglobulin test ,free antibody and antibody releasing test ( hemolysis three tests ) and measurement of serum total bili-rubin for hemolytic disease of newborn ( HDN) .Methods Hemolysis three tests were carried out with micro column gel technology in 212 blood samples from neonatal jaundice ( derived from the O type mothers ) and positive rate differ-ences in hemolysis three tests were compared .Patients were divided into 4 groups by the antibody results in hemolysis three tests:group 1[direct antiglobulin test ( +),free antibody test ( +) and antibody releasing test ( +)],group 2[(direct antiglobulin test(+),free antibody test(-) and antibody releasing test(+)],group 3[direct antiglobu-lin test(-),free antibody test(+) and antibody releasing test(+),and group 4[direct antiglobulin test( -),free antibody test (-) and antibody releasing test (+) ] .Constituent ratio was compared .The total bilirubin in serum from four groups were determined by automatic biochemical analyzer and compared .Results The positive rates of direct antiglobulin test,free antibody test and antibody release test were 18.4%(39/212),33.5%(71/212) and 39.6%(84/212) respectively.The positive rate had no significant difference between antibody release test and free antibody test(χ2 =1.70,P>0.05),but compared with direct antiglobulin test ,the differences were significant (χ2 =23.18, 12.58,all P<0.05).Serum total bilirubin levels in four groups were significantly increased ,but had no significant differences(F=1.369,P>0.05).Conclusion Combined detection of serum total bilirubin and serum hemolysis three tests can be used as the early diagnosis and patients condition of HDN ,and contribute to the early control of neo-natal hemolytic increase ,reduce complications and sequelae .