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The page below is a sample from the LabCE course Hemolytic Disease of the Fetus and Newborn. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Newborn Serologic Testing Protocols

Protocols for testing newborns vary internationally and within countries. The table below summarizes some of the more common protocols.


Typical Newborn Testing Protocol


Mother is D-negative with no unexpected antibodies

Newborn is tested at delivery for:

  • ABO and Rh
  • Test for weak D (mandatory) if initial Rh typing appears to be D-negative
  • Direct antiglobulin test (DAT)*

A positive DAT does not always mean that the newborn has clinically significant hemolysis.

  • A positive DAT commonly occurs due to ABO incompatibility, yet infants seldom require treatment.
  • Infants born to mothers who received antenatal RhIg sometimes have a positive DAT that does not cause clinically relevant hemolysis.

Mother is Rh positive and a blood group other than group O

  • Routine testing not performed
  • Cord blood retained for a specified period of time (e.g., seven days) in the event that the mother has an unexpected antibody at delivery or the newborn develops signs of red cell hemolysis.
  • Routine testing would result in many positive DATs due to ABO incompatibility- not clinically significant.

Mother is group O Rh positive

Newborn is tested- especially important if women and their infants are discharged within 24 hours since hyperbilirubinemia due to ABO HDFN may develop later.
  • Optional only if there is appropriate surveillance and risk assessment before discharge and provided there is follow-up (American Academy of Pediatrics).

*Policies for DAT testing of newborns whose mothers have received antenatal RhIg vary internationally. For example, the British Committee for Standards in Haematology guidelines state that a DAT should not be performed on cord blood routinely since in some cases it may be positive due to antenatal RhIg prophylaxis. A DAT is recommended only if HDFN is suspected because of a low cord blood hemoglobin or the presence of unexpected maternal antibodies. However in North America, DATs are always performed on infants born to Rh negative mothers who are RhIg candidates.