title: Strategies for rare-event detection: an approach for automated fetal cell detection in maternal blood.
abstract: This article explores the feasibility of the use of automated microscopy and image analysis to detect the presence of rare fetal nucleated red blood cells (NRBCs) circulating in maternal blood. The rationales for enrichment and for automated image analysis for "rare-event" detection are reviewed. We also describe the application of automated image analysis to 42 maternal blood samples, using a protocol consisting of one-step enrichment followed by immunocytochemical staining for fetal hemoglobin (HbF) and FISH for X- and Y-chromosomal sequences. Automated image analysis consisted of multimode microscopy and subsequent visual evaluation of image memories containing the selected objects. The FISH results were compared with the results of conventional karyotyping of the chorionic villi. By use of manual screening, 43% of the slides were found to be positive (>=1 NRBC), with a mean number of 11 NRBCs (range 1-40). By automated microscopy, 52% were positive, with on average 17 NRBCs (range 1-111). There was a good correlation between both manual and automated screening, but the NRBC yield from automated image analysis was found to be superior to that from manual screening (P=.0443), particularly when the NRBC count was >15. Seven (64%) of 11 XY fetuses were correctly diagnosed by FISH analysis of automatically detected cells, and all discrepancies were restricted to the lower cell-count range. We believe that automated microscopy and image analysis reduce the screening workload, are more sensitive than manual evaluation, and can be used to detect rare HbF-containing NRBCs in maternal blood.
authors.affiliation: Department of Cytochemistry, Leiden University Medical Center, The Netherlands. j.c.oosterwijk@med.run.nl
authors.forename: J C
authors.initials: JC
authors.lastname: Oosterwijk
authors.forename: C F
authors.initials: CF
authors.lastname: Knepflé
authors.forename: W E
authors.initials: WE
authors.lastname: Mesker
authors.forename: H
authors.initials: H
authors.lastname: Vrolijk
authors.forename: W C
authors.initials: WC
authors.lastname: Sloos
authors.forename: H
authors.initials: H
authors.lastname: Pattenier
authors.forename: I
authors.initials: I
authors.lastname: Ravkin
authors.forename: G J
authors.initials: GJ
authors.lastname: van Ommen
authors.forename: H H
authors.initials: HH
authors.lastname: Kanhai
authors.forename: H J
authors.initials: HJ
authors.lastname: Tanke
mesh_terms: D001331:Automation; D002469:Cell Separation; D004900:Erythroblasts / Q000166:cytology*; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005319:Fetal Hemoglobin / Q000032:analysis; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008297:Male; D008853:Microscopy / Q000379:methods*; D008856:Microscopy, Fluorescence; D011247:Pregnancy / Q000097:blood*; D012680:Sensitivity and Specificity; D012730:Sex Chromosomes; D013194:Staining and Labeling
chemical_list: D005319:Fetal Hemoglobin