[Fetal nucleated erythrocytes in the peripheral circulation of pregnant women as a noninvasive method of fetal sex determination].
OBJECTIVE
Determination of the sex of the foetus from enriched nuclear red blood cells (NRBC) circulating in maternal blood during pregnancy.


METHODS
NRBC were enriched from 13-28 ml peripheral blood of 32 pregnant women using double MACS procedure. NRBCs were enriched by magnetic activated cell sorting using anti-CD71 (transferrin receptor) monoclonal antibodies. Unwanted leukocytes were depleted using monoclonal antibodies against CD14 and CD45. The sex of the foetus was analysed by using dual-colour FISH with X and Y specific probes. The experimental results obtained from the noninvasive procedure were compared to karyotype obtained from amniocentesis or chorionic villus sampling.


RESULTS
In 15 out of 17 male foetuses we could identify one X and one Y signal. In another 15 pregnant women carrying female foetuses two X signals were observed.


CONCLUSIONS
NRBC circulating in blood of pregnant women can be used as an alternative source for determination of the sex of the foetus with a risk of false negative results (2/17, 12%). The problem of false negative results can be solved by using more sophisticated methods of enrichment and preparedness of the slides for FISH analysis.
Hromadíková
I
I
II. dĕtská klinika 2. LF UK FN Motol, Praha.
Karamanov
S
S
Mrstinová
M
M
Stechová
K
K
Kofer
J
J
Houbová
B
B
Macek
M
M
D015342:DNA Probes; D004900:Erythroblasts; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D011247:Pregnancy / Q000097:blood*; D012732:Sex Determination Analysis / Q000379:methods*
D015342:DNA Probes