Isolation of epsilon-haemoglobin-chain positive fetal cells with micromanipulation for prenatal diagnosis.
OBJECTIVE
The isolation and analysis of fetal cells in maternal blood during pregnancy is under investigation as a means of noninvasive prenatal diagnosis. The aim of our study was to detect fetal gender from maternal peripherial blood samples during pregnancy with the detection and analysis of epsilon-haemoglobin-chain positive fetal nucleated red blood cells (NRBCs) collected by a micromanipulator. Here we report our first results.


METHODS
We obtained maternal blood from 14 singleton pregnancies. After a double density gradient separation, magnetic activated cell sorting was performed by positive selection for nucleated red blood cells with anti-CD71. With the help of this enrichment step, followed by immunophenotyping with an anti-haemoglobin-epsilon monoclonal antibody, the isolation of the epsilon haemoglobin chain positive cells with micromanipulation could be done. We performed single cell fluorescent PCR analysis of these cells; we used primers for the amelogenin gene to detect fetal gender. We compared our findings with the results of amniocentesis.


RESULTS
Fetal gender was successfully determined in 11 out of 14 cases; among them, in 2 cases with Klinefelter syndrome (47,XXY).


CONCLUSIONS
The results of our study suggest that micromanipulation and QF-PCR analysis of anti-haemoglobin-epsilon fluorescent antibody stained fetal cells from maternal blood can be useful in prenatal diagnosis to detect fetal gender and promising to be improved to detect chromosomal abnormalities.
Nagy
Gyula Richárd
GR
Department of Obstetrics and Gynecology, Semmelweis University, Faculty of Medicine, Budapest, Hungary. nagygyr@noi1.sote.hu
Bán
Zoltán
Z
Sipos
Ferenc
F
Beke
Artúr
A
Papp
Csaba
C
Papp
Zoltán
Z
D015703:Antigens, CD / Q000276:immunology*; D004900:Erythroblasts / Q000166:cytology*; D005260:Female; D005319:Fetal Hemoglobin / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D005914:Globins / Q000032:analysis*; D006801:Humans; D008297:Male; D008846:Micromanipulation; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis; D019849:Sex Determination Processes
D015703:Antigens, CD; D005914:Globins; D005319:Fetal Hemoglobin