Laboratory methods for Rh immunoprophylaxis: a review

The recommended dose of Rh immune globulin for postpartum Rh immunoprophylaxis is based on an estimation of the volume of the fetomaternal hemorrhage, if any, measured as the percent of fetal RBCs in a sample of the D– mother's blood. Laboratory methods for distinguishing fetal from maternal RBCs have been based on their different blood types (D+ versus D–) or predominant hemoglobin content (hemoglobin F versus hemoglobin A). We conducted a review of the medical literature describing laboratory methods for detecting and quantifying fetal RBCs in maternal blood samples. We also used data collected for the College of American Pathologists Fetal RBC Detection Surveys to determine which laboratory methods are used currently in hospitals in the United States. The rosette screen is used widely for identifying D– mothers who may require additional doses of Rh immune globulin for postpartum immunoprophylaxis. As the rosette screen targets the D antigen, it is not suitable for detecting a fetomaternal hemorrhage in D+ mothers or when the D type of the fetus or newborn is D– or unknown. The acid-elution (Kleihauer- Betke) assay is a sensitive laboratory method for quantifying a fetomaternal hemorrhage, but it is tedious, often inaccurate, and difficult to reproduce. Flow cytometry, using anti-D or anti-hemoglobin F reagents, offers a more precise quantification of fetal RBCs in maternal blood. However, flow cytometry services for this function are available in relatively few hospital laboratories in the United States because of logistic and fiscal impediments.