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To assess the utility of measuring direct bilirubin (DB) during the first 72 h of life in infants admitted to the neonatal intensive care unit (NICU).Infants born between May 2006 and June 2013, and admitted to the NICU were included. Abnormal DB was defined as: DB level≥1 mg dl-1 with a corresponding TB of ≤5 mg dl-1 or DB level >20% of the corresponding TB>5 mg dl-1.The DB levels were measured in 3715 infants during the first 72 h of life. Sixty-three infants (1.7%) had abnormal DB. In a number of infants with potentially treatable diseases (biliary atresia and choledochal cyst), the only abnormal finding was a mildly elevated DB (1 to 2 mg dl-1) during the first 3 days of life. In 22 infants (35%), the cause for high DB was unknown (16%) or not investigated (19%).Routine measurement of DB in neonates admitted to NICU may be helpful in identifying potentially treatable causes of cholestasis.