Document Detail


Use of the Kleihauer test to detect fetal erythroblasts in the maternal circulation.
MedLine Citation:
PMID:  11241536     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Non-invasive prenatal diagnosis of aneuploidies on fetal nucleated erythrocytes present in the maternal circulation is hampered by the extremely small cell number of uncertain origin (70% of erythroblasts circulating during pregnancy have a maternal origin). Therefore, a method allowing selection of the fetal cells among the maternal cells is indispensable after the erythroblast enrichment step. In the present study, after an erythroblast enrichment step on a ficoll gradient followed by a positive immuno-magnetic selection with anti-CD71 or anti-GPA antibodies, a rapid, simple and direct chemical staining method adapted from the classical Kleihauer test was developed to select fetal cells. Precise differentiation between fetal and maternal erythroblasts is based on the constitutional difference between fetal and adult haemoglobin (Hb). The fetal cells appear with an intense pink cytoplasmic staining while maternal cells with adult haemoglobin are colourless. Preservation of the cytoplasmic integrity allows one to distinguish morphological characteristics and to visualize simultaneously nuclear hybridization signal by FISH (fluorescent in situ hybridization). This approach was tested by FISH analysis using dual-colour X- and Y-specific DNA probes on blood samples from 15 pregnant women, with the results being compared to cytogenetic or sonographic sex determination. For 12 pregnancies fetal sex was determined successfully (5 XY/7 XX), in two cases in situ hybridization failed, and in one case no fetal erythroblast was observed after the Kleihauer test. The selection method was applied to a pregnancy at risk for cystic fibrosis (CF). After a Kleihauer test, fetal erythroblasts were collected by microdissection, whole genomic DNA was amplified by primer extension pre-amplification (PEP) followed by a nested CF PCR. The fetal genotype was successfully characterized and confirmed by conventional prenatal diagnosis.
Authors:
V Martel-Petit; C Petit; M Marchand; A Fleurentin; B Fontaine; A Miton; P Lemarie; C Philippe; P Jonveaux
Related Documents :
1259926 - The kenya form of hereditary persistence of fetal haemoglobin: structural studies and e...
17893646 - Expression of sp1 and klf6 in the developing human cornea.
1238566 - Ultrastructural studies of the rabbit placenta in the last third of gestation.
11726596 - Ultrastructure of epithelial plaque formation and stromal cell transformation by post-o...
16415966 - Chromosomal abnormalities and adverse pregnancy outcome with maternal serum second trim...
17940926 - Movement disorders in pregnancy.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Prenatal diagnosis     Volume:  21     ISSN:  0197-3851     ISO Abbreviation:  Prenat. Diagn.     Publication Date:  2001 Feb 
Date Detail:
Created Date:  2001-03-12     Completed Date:  2001-06-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8106540     Medline TA:  Prenat Diagn     Country:  England    
Other Details:
Languages:  eng     Pagination:  106-11     Citation Subset:  IM    
Copyright Information:
Copyright 2001 John Wiley & Sons, Ltd.
Affiliation:
Laboratoire de Génétique Médicale, CHU Nancy, France.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Antibodies
Antigens, CD / immunology
Antigens, Differentiation, B-Lymphocyte / immunology
Cell Nucleus / ultrastructure
Coloring Agents
Cystic Fibrosis / blood,  diagnosis,  genetics
Cytoplasm / chemistry
Erythroblasts* / chemistry,  ultrastructure
Female
Fetal Blood / cytology*
Fetal Hemoglobin / analysis
Genotype
Gestational Age
Glycophorin / immunology
Humans
Immunomagnetic Separation
In Situ Hybridization, Fluorescence
Male
Polymerase Chain Reaction
Pregnancy
Receptors, Transferrin
Chemical
Reg. No./Substance:
0/Antibodies; 0/Antigens, CD; 0/Antigens, Differentiation, B-Lymphocyte; 0/CD71 antigen; 0/Coloring Agents; 0/Glycophorin; 0/Receptors, Transferrin; 9034-63-3/Fetal Hemoglobin

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


Previous Document:  Prenatal diagnosis of mosaic ring chromosome 13 with anencephaly.
Next Document:  Prenatal measurement of testicular diameter by ultrasonography: development of fetal male gender and...