A combined flow cytometry-based method for fetomaternal hemorrhage and maternal D

Darcie J. Radel, Connie S. Penz, Allan B Dietz, Dennis A. Gastineau

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

BACKGROUND: The fetomaternal bleed assay by flow cytometry is a semiquantitative test used to determine the volume of fetomaternal hemorrhage (FMH). We have enhanced this method by combining the fetomaternal bleed assay with a D antibody in the same tube to determine the maternal D as well as the need for Rh immune globulin (RhIG) administration. STUDY DESIGN AND METHODS: The performance of the FMB/D assay was compared against the Kleihauer-Betke (KB) acid elution method for the quantitation of fetal bleed and tube method for the maternal D determination. Peripheral blood cells from the pregnant women were fixed and permeabilized, incubated with monoclonal antibodies directed against fetal hemoglobin (Hb F) and D, and analyzed by flow cytometry. The relative concentration of Hb F-containing cells to adult cells was used to determine the volume of the FMH. In addition, the cells were identified as D+ or D-. RESULTS: D typing of nonpregnant adult donors matched in all cases with the tube method with a mean value of 0.01% Hb F-positive cells (n = 25). D typing of pregnant or postpartum samples matched in all cases (n = 100), and the RhIG dose calculated by the percentage of Hb F for each method matched 97 percent (n = 36; p = 0.324). The one discrepant sample was consistent with the acknowledged overestimation when using the KB method. CONCLUSION: The highly precise anti-Hb F flow cytometric method was combined with maternal D determination by anti-D. This method correlates well with standard assays eliminating the need for additional testing without sacrificing clinical information.

Original languageEnglish (US)
Pages (from-to)1886-1891
Number of pages6
JournalTransfusion
Volume48
Issue number9
DOIs
StatePublished - Sep 2008

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Fetomaternal Transfusion
Flow Cytometry
Mothers
Rho(D) Immune Globulin
Fetal Hemoglobin
Postpartum Period
Pregnant Women
Blood Cells
Monoclonal Antibodies
Tissue Donors

ASJC Scopus subject areas

  • Hematology
  • Immunology

Cite this

A combined flow cytometry-based method for fetomaternal hemorrhage and maternal D. / Radel, Darcie J.; Penz, Connie S.; Dietz, Allan B; Gastineau, Dennis A.

In: Transfusion, Vol. 48, No. 9, 09.2008, p. 1886-1891.

Research output: Contribution to journalArticle

Radel, Darcie J. ; Penz, Connie S. ; Dietz, Allan B ; Gastineau, Dennis A. / A combined flow cytometry-based method for fetomaternal hemorrhage and maternal D. In: Transfusion. 2008 ; Vol. 48, No. 9. pp. 1886-1891.
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