PMID- 11468149
OWN - NLM
STAT- MEDLINE
DCOM- 20010913
LR  - 20210216
IS  - 0006-4971 (Print)
IS  - 0006-4971 (Linking)
VI  - 98
IP  - 3
DP  - 2001 Aug 1
TI  - Simultaneous fetal cell identification and diagnosis by epsilon-globin chain 
      immunophenotyping and chromosomal fluorescence in situ hybridization.
PG  - 554-7
AB  - Isolating fetal erythroblasts from maternal blood offers a promising noninvasive 
      alternative for prenatal diagnosis. The current immunoenzymatic methods of 
      identifying fetal cells from background maternal cells postenrichment by labeling 
      gamma-globin are problematic. They are nonspecific because maternal cells may 
      produce gamma-globin, give poor hybridization efficiencies with chromosomal 
      fluorescence in situ hybridization (FISH), and do not permit simultaneous 
      visualization of the fetal cell identifier and the FISH signal. We describe a 
      novel technique that allows simultaneous visualization of fetal erythroblast 
      morphology, chromosomal FISH, and epsilon-globin labeled with AMCA 
      (7-amino-4-methylcoumarin-3-acetic acid). AMCA was chosen as the fluorescent 
      label to circumvent the problem of heme autofluorescence because the mean 
      difference in relative fluorescence intensity between fetal erythroblasts stained 
      positive for antiglobin antibody and autofluorescence of unstained cells was 
      greater with AMCA (mean 43.2; 95% confidence interval [CI], 34.6-51.9; SD = 14.0) 
      as the reporting label compared with fluorescein isothiocyanate (mean 24.2; 95% 
      CI, 16.4-31.9; SD = 12.4) or phycoerythrin (mean 9.8; 95% CI, 4.8-14.8; SD = 
      8.0). Median FISH hybridization efficiency was 97%, comparable to the 98% (n = 5 
      paired samples) using Carnoy fixative. One epsilon-positive fetal erythroblast 
      was identified among 10(5) maternal nucleated cells in 6 paired mixture 
      experiments of fetal erythroblasts in maternal blood (P <.001). Male 
      epsilon-positive fetal erythroblasts were clearly distinguishable from adult 
      female epsilon-negative erythroblasts, with no false positives (n = 1000). The 
      frequency of fetal erythroblasts expressing epsilon-globin declines linearly from 
      7 to 14 weeks' gestation (y = -15.8 x + 230.8; R(2) = 0.8; P <.001). We describe 
      a rapid and accurate method to detect simultaneously fetal erythroblast 
      morphology, intracytoplasmic epsilon-globin, and nuclear FISH. (Blood. 
      2001;98:554-557)
FAU - Choolani, M
AU  - Choolani M
AD  - Department of Maternal and Fetal Medicine, Division of Paediatrics, Institute of 
      Reproductive and Developmental Biology, Imperial College School of Medicine, 
      Hammersmith Hospital Campus, Du Cane Rd., London W12 0NN, United Kingdom. 
      mchoolani@cwcom.net
FAU - O'Donnell, H
AU  - O'Donnell H
FAU - Campagnoli, C
AU  - Campagnoli C
FAU - Kumar, S
AU  - Kumar S
FAU - Roberts, I
AU  - Roberts I
FAU - Bennett, P R
AU  - Bennett PR
FAU - Fisk, N M
AU  - Fisk NM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Blood
JT  - Blood
JID - 7603509
RN  - 0 (Biomarkers)
RN  - 9004-22-2 (Globins)
SB  - IM
MH  - Adult
MH  - Biomarkers/analysis
MH  - Cytogenetic Analysis
MH  - Erythroblasts/chemistry/*cytology/ultrastructure
MH  - Female
MH  - Fetal Blood/*cytology
MH  - Gestational Age
MH  - Globins/*analysis
MH  - Humans
MH  - Immunohistochemistry
MH  - Immunophenotyping
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - Pregnancy
MH  - Prenatal Diagnosis/*methods
EDAT- 2001/07/27 10:00
MHDA- 2001/09/14 10:01
CRDT- 2001/07/27 10:00
PHST- 2001/07/27 10:00 [pubmed]
PHST- 2001/09/14 10:01 [medline]
PHST- 2001/07/27 10:00 [entrez]
AID - S0006-4971(20)48050-4 [pii]
AID - 10.1182/blood.v98.3.554 [doi]
PST - ppublish
SO  - Blood. 2001 Aug 1;98(3):554-7. doi: 10.1182/blood.v98.3.554.