title: Quantification of fetomaternal hemorrhage: a comparative study of the manual and automated microscopic Kleihauer-Betke tests and flow cytometry in clinical samples.
abstract: OBJECTIVE
  The purpose of this study was to evaluate the quantification of fetomaternal hemorrhage by the manual and automated microscopic analysis of Kleihauer-Betke stained slides and by flow cytometry.
  
  
  METHODS
  Blood smears were stained and evaluated manually according to the Kleihauer-Betke test. The same slides were used for automated microscopy. In addition, blood flow cytometry was performed by anti-hemaglobin F immunostaining.
  
  
  RESULTS
  Fetomaternal hemorrhage >0.1% was detected in 4 patients by manual and automated Kleihauer-Betke test and by blood flow cytometry. Fetomaternal hemorrhage was absent according to all 3 methods in 13 patients; fetomaternal hemorrhage<0.1% was detected in 27 patients by either manual or automated Kleihauer-Betke test or both. Moderate agreement was observed between the manual and automated Kleihauer-Betke test (weighted kappa, 0.56; 95% CI, 0.33-0.78). Agreement between the manual Kleihauer-Betke test and blood flow cytometry was fair (weighted kappa, 0.40; 95% CI, 0.15-0.66).
  
  
  CONCLUSIONS
  Automated microscopic detection of fetal blood cells in clinical samples provides accurate quantification that is comparable to the manual Kleihauer-Betke test in both small and large fetomaternal hemorrhage. Blood flow cytometry is capable only of quantifying fetomaternal hemorrhage of >0.1%.
authors.affiliation: Department of Obstetrics, Laboratory for Cytochemistry and Cytometry, Department of Molecular Cell Biology, Department of Hematology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
authors.forename: Denise M
authors.initials: DM
authors.lastname: Pelikan
authors.forename: Sicco A
authors.initials: SA
authors.lastname: Scherjon
authors.forename: Wilma E
authors.initials: WE
authors.lastname: Mesker
authors.forename: Godelieve M
authors.initials: GM
authors.lastname: de Groot-Swings
authors.forename: Geeske G
authors.initials: GG
authors.lastname: Brouwer-Mandema
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Tanke
authors.forename: Humphrey H
authors.initials: HH
authors.lastname: Kanhai
mesh_terms: D004911:Erythrocyte Volume; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005331:Fetomaternal Transfusion / Q000175:diagnosis* / Q000209:etiology; D005434:Flow Cytometry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008853:Microscopy / Q000379:methods; D011247:Pregnancy; D018566:Pregnancy, High-Risk; D012203:Rh Isoimmunization / Q000150:complications*