Analysis of paternal alleles in nucleated red blood cells enriched from maternal blood

The purpose of our study was to identify paternal alleles in NRBC enriched from maternal peripheral blood for detection of the presence of foetal cell s in the maternal circulation and to establish a reliable non-invasive meth od which should allow following genetic testing. For enrichment of foetal c ells from peripheral maternal blood we combined Ficoll-Paque density gradie nt centrifugation and MACS, Maternal leukocytes were firstly depleted using anti-CD14 and anti-CD45 microbeads. NRBC were sorted from the CD14(-)/CD45 (-) fraction by positive selection using CD71 microbeads, Paternal alleles in the CD14(-)/CD45(-)/CD71(+) fraction were indicated by the PCR method us ing HLA (DRB1, DQB1, DQA1) and Polymarker System (LDLR, GYPA, HBGG, D7S8, G C) as genetic markers. Different paternal alleles of studied 8 loci were de tected in 13 out of 19 samples of cells enriched from maternal peripheral b lood between the 13(th) and 36(th) week of gestation. Our results demonstra te that foetal cells enriched from maternal peripheral blood may be used as a source of foetal DNA for prenatal diagnosis, paternity testing and other application.