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Fetal nucleated erythrocytes (NRBCS) in chorionic villus sample supernatant fluids: an additional source of fetal material for karyotype confirmation

Fetal erythrocytes leak from the fetal capillaries at the time of chorionic villus removal. The purpose of this study was to determine if fetal nucleated erythrocytes (NRBCs) could be isolated from the chorionic villus sampling (CVS) supernatant fluid and used as an additional source of fetal material in order to confirm the fetal karyotype in cases of CVS mosaicism. One hundred CVS supernatant fluids were studied by simultaneous immunophenotyping, using a mouse antifetal haemoglobin antibody, UCHγ, combined with fluorescent in situ hybridization (FISH) analysis using X‐ and Y‐specific DNA probes. A chromosome 18 probe was also used in the case of a known male fetus with trisomy 18. Fetal haemoglobin (HbF)‐positive cells were identified in 73 supernatant fluids and HbF‐positive nucleated cells were present in 60 samples. The number of cells detected per sample showed great variation among the individual samples. FISH analysis was performed in 41 cases. FISH prediction of the fetal gender was concordant with the CVS karyotype in all cases, and the fetal trisomy 18 was correctly verified. In five cases in which Y sequences were detected, a small number of HbF‐positive cells with two X signals were also identified; interestingly, in three of the five cases, the mother was a β‐thalassaemia carrier. This technique can be used as a quick and accurate method for the immediate verification of CVS results in cases of mosaicism, thus avoiding second‐trimester amniocentesis. © 1997 John Wiley & Sons, Ltd.