Document Detail


Simultaneous fetal cell identification and diagnosis by epsilon-globin chain immunophenotyping and chromosomal fluorescence in situ hybridization.
MedLine Citation:
PMID:  11468149     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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)
Authors:
M Choolani; H O'Donnell; C Campagnoli; S Kumar; I Roberts; P R Bennett; N M Fisk
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Blood     Volume:  98     ISSN:  0006-4971     ISO Abbreviation:  Blood     Publication Date:  2001 Aug 
Date Detail:
Created Date:  2001-07-24     Completed Date:  2001-09-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7603509     Medline TA:  Blood     Country:  United States    
Other Details:
Languages:  eng     Pagination:  554-7     Citation Subset:  AIM; IM    
Affiliation:
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
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MeSH Terms
Descriptor/Qualifier:
Adult
Biological Markers / analysis
Cytogenetic Analysis
Erythroblasts / chemistry,  cytology*,  ultrastructure
Female
Fetal Blood / cytology*
Gestational Age
Globins / analysis*
Humans
Immunohistochemistry
Immunophenotyping
In Situ Hybridization, Fluorescence
Male
Pregnancy
Prenatal Diagnosis / methods*
Chemical
Reg. No./Substance:
0/Biological Markers; 9004-22-2/Globins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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