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Hemolytic disease of the newborn (HDN) is a potentially fatal alloimmune condition where fetal red blood cells are destroyed by transplacentally acquired maternal antibodies. Rh disease (RhD) is the most common offending paternal antigen.
Reproduced with permission from Humana Press. Liley Chart. (Used with permission from Grenache, 2004, 231)
Reproduced with permission from Humana Press. Queenan Chart. (Used with permission from Grenache, 2004, 235)
Tests generally appear in the order most useful for common clinical situations
| Test name: Antigen Testing, Rh Phenotype |
| ARUP #: 0013019 |
| Methodology: Hemagglutination |
| Use: Maternal testing to assess Rh status Fetal testing after pregnancy Antigen testing for D, C, E, c, e |
| Test name: Amniotic Bilirubin Scan |
| ARUP #: 0080276 |
| Methodology: Quantitative Spectrophotometry (Delta OD 450 nm) |
| Use: Assess alloimmune hemolytic disease of the fetus Follow progression of disease to determine need for fetal transfusion or early delivery |
| Limitations: Bloody amniotic fluid compromises the accuracy of results |
| Follow-up: Evaluate in conjunction with fetal Rh genotyping Ultrasound measurement of the middle cerebral artery blood velocity can estimate fetal anemia |
| Test name: RhD Antigen (RhD) Genotyping |
| ARUP #: 0051368 |
| Methodology: Polymerase Chain Reaction/Fluorescence Monitoring |
| Use: Paternal testing to determine RHD heterozygosity or homozygosity in a phenotypically positive individual when the mother has clinically significant alloantibody If the father is determined to be homozygous for the RHD allele, offspring can be assumed to be RhD positive, negating the need for fetal RhD testing Fetal testing when the mother has clinically significant alloantibody and the father is either heterozygous for RHD or not available for testing |
| Limitations: Bloody amniotic fluid specimens may give false negative results from maternal cell contamination Specificity may be compromised by mutations in primer sites or those outside the RHCE exons examined Fetuses predicted to be unaffected should continue to be monitored by noninvasive means Most rare mutations in the RHD gene (ie, missense, nonsense, insertions, gene fusion, or small deletions) will not be detected by this assay, and the sample may be misinterpreted as being Rh-positive Clinical sensitivity greater than or equal to 98% |
| Test name: RhCc Antigen (RHCE) Genotyping |
| ARUP #: 0050421 |
| Methodology: Polymerase Chain Reaction/Fluorescent Monitoring |
| Use: Fetal testing when the mother has a clinically significant alloantibody level and the father is phenotypically positive for the RHCE gene encoding the corresponding antigen |
| Limitations: Bloody amniotic fluid specimens may give false-negative results because of maternal cell contamination Specificity may be compromised by mutations in primer sites or those outside the RHCE exons examined Fetuses predicted to be unaffected should continue to be monitored by noninvasive means Individuals with weak or no expression of the Cc/Ee antigens may result from RHCE gene alterations such as RHCE-D-CE gene hybrids; other hybrids allow for expression of the C, c, or e antigens on the RHD allele Genotyping may result in false-negative RhC, Rhc, or Rhe predictions due to RHCE-D-CE fusion genes Clinical sensitivity: unknown |
| Test name: RhEe Antigen (RHCE) Genotyping |
| ARUP #: 0050423 |
| Methodology: Polymerase Chain Reaction/Fluorescent Monitoring |
| Use: Fetal testing when the mother has a clinically significant alloantibody level and the father of the pregnancy is phenotypically positive for the RHCE gene encoding the corresponding antigen |
| Limitations: Bloody amniotic fluid specimens may give false-negative results from maternal cell contamination Specificity may be compromised by mutations in primer sites or those outside the RHCE exons examined Fetuses predicted to be unaffected should continue to be monitored by noninvasive means Weak or no expression of Cc/Ee antigens may result from RHCE gene alterations such as RHCE-D-CE hybrids; other hybrids allow for expression of the C, c, or e antigens on the RHD allele Genotyping may result in false-negative RhC, Rhc, or Rhe predictions due to RHCE-D-CE fusion genes Clinical sensitivity unknown |
| Test name: Kell K/k Antigen (KEL) Genotyping |
| ARUP #: 0051644 |
| Methodology: Polymerase Chain Reaction/Fluorescence Monitoring |
| Use: Fetal testing when the mother has clinically significant alloantibody and the father of the pregnancy is either heterozygous for KEL1 or is not available for testing Paternal testing in a KEL1 RBC antigen-positive individual to determine KEL1 heterozygosity or homozygosity when the mother is KEL1-negative by RBC antigen typing |
| Limitations: KEL antigens other than KEL1/KEL2 are not evaluated by this assay Bloody amniotic fluid samples may give false-negative results due to maternal cell contamination If the father is determined to be homozygous for the KEL1, all of his offspring can be assumed to be KEL1-positive, negating the need for fetal KEL1 testing |
| Test name: Fetal Hemoglobin Determination for Fetomaternal Hemorrhage |
| ARUP #: 2001743 |
| Methodology: Quantitative Flow Cytometry |
| Use: Used to detect and quantify the extent of fetomaternal hemorrhage in pregnant or post-partum women and assess the need for Rh immune globulin (eg RhoGAM®) for fetomaternal hemorrhage |
| Test name: Hemoglobin F |
| ARUP #: 0081348 |
| Methodology: High Performance Liquid Chromatography |
| Comments: |
| Test name: Kleihauer-Betke Stain for Fetal Hemoglobin |
| ARUP #: 0040105 |
| Methodology: Semi-Quantitative Acid Elution Eosin Stain/Microscopy |
| Comments: Do not use to detect fetomaternal hemorrhage |
| Test name: ABO-Rh Type |
| ARUP #: 0010025 |
| Methodology: Hemagglutination |
| Comments: |
| Test name: Bilirubin, Direct, Serum or Plasma |
| ARUP #: 0020033 |
| Methodology: Quantitative Spectrophotometry |
| Comments: |