PMID- 12533817
OWN - NLM
STAT- MEDLINE
DCOM- 20030702
LR  - 20101118
IS  - 0197-3851 (Print)
IS  - 0197-3851 (Linking)
VI  - 23
IP  - 1
DP  - 2003 Jan
TI  - A simple and sensitive erythroblast scoring system to identify fetal cells in 
      maternal blood.
PG  - 68-73
AB  - OBJECTIVES: Nucleated red blood cells (NRBCs) of fetal origin appear to have 
      distinguishable characteristics from that of maternal NRBCs in both nuclear 
      morphology and properties of hemoglobin staining. However, these differences have 
      yet to be quantified. Our aim was to develop an erythroblast scoring system using 
      four distinct phenotypic parameters (nuclear roundness, nuclear morphology, gamma 
      hemoglobin staining intensity, and peripheral brightness of the stained 
      cytoplasm) to address this issue. METHODS: NRBCs were isolated from four maternal 
      blood samples by density gradient separation, CD15/45 depletion, and gamma 
      hemoglobin positive selection after elective termination of a trisomy 21 male 
      fetus (47,XY,+21). All cells were deposited onto microscope slides and every NRBC 
      was analyzed according to the scoring system. Each of the four individual 
      parameters was given a value from 0 to 3 points and a combined score was obtained 
      for each cell (range 0-12). Fluorescence in situ hybridization (FISH) using X- 
      and Y-specific probes was performed to determine, on the basis of interphase 
      karyotype, whether the cell was maternal or fetal. RESULTS: The majority of 
      maternal NRBCs were found to have a combined score of 6 or less (103/117) and the 
      majority of fetal NRBCs were found to have a score of 7 or greater (43/53). The 
      proportion of cells that were identified correctly as fetal increased with each 
      ascending category of combined score. For example, 5.7% of NRBCs with a combined 
      score of 5 points or less were found to be fetal, whereas 19.2% of NRBCs with a 
      combined score of 6 points were fetal. At combined scores of 11 and 12 points, 
      100% of NRBCs were found to be fetal. CONCLUSION: Fetal NRBCs have characteristic 
      morphology and a gamma hemoglobin staining appearance that makes them 
      distinguishable from maternal NRBCs. The scoring system presented here is a 
      simple and sensitive method to distinguish fetal NRBCs from adult cells in 
      maternal blood. This system may have clinical utility for noninvasive prenatal 
      diagnosis as well as applications for basic research into the developmental 
      biology of NRBCs. In addition, these defined parameters may serve as 
      computational classifiers for the automated detection of fetal cells in maternal 
      blood.
CI  - Copyright 2003 John Wiley & Sons, Ltd.
FAU - Cha, Donghyun
AU  - Cha D
AD  - Division of Genetics, Departments of Obstetrics and Gynecology and Pediatrics, 
      Tufts-New England Medical Center, Boston, MA 02111, USA.
FAU - Hogan, Brad
AU  - Hogan B
FAU - Bohmer, Ralph M
AU  - Bohmer RM
FAU - Bianchi, Diana W
AU  - Bianchi DW
FAU - Johnson, Kirby L
AU  - Johnson KL
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Prenat Diagn
JT  - Prenatal diagnosis
JID - 8106540
RN  - 9034-63-3 (Fetal Hemoglobin)
SB  - IM
MH  - Adult
MH  - Cell Nucleus/classification
MH  - Cell Separation
MH  - Cytoplasm/chemistry/classification
MH  - Erythroblasts/chemistry/*cytology
MH  - Female
MH  - Fetal Blood/chemistry/*cytology
MH  - Fetal Hemoglobin/*analysis
MH  - Fetomaternal Transfusion/*blood
MH  - Flow Cytometry
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Leukocytes, Mononuclear/cytology/metabolism
MH  - Male
MH  - Micromanipulation
MH  - Pregnancy/blood
MH  - Prenatal Diagnosis/*methods
MH  - Sensitivity and Specificity
MH  - Sex Determination Analysis
EDAT- 2003/01/21 04:00
MHDA- 2003/07/03 05:00
CRDT- 2003/01/21 04:00
PHST- 2003/01/21 04:00 [pubmed]
PHST- 2003/07/03 05:00 [medline]
PHST- 2003/01/21 04:00 [entrez]
AID - 10.1002/pd.515 [doi]
PST - ppublish
SO  - Prenat Diagn. 2003 Jan;23(1):68-73. doi: 10.1002/pd.515.