Fol. Biol. 2001, 47, 36-39
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 cells in the maternal circulation and to establish a reliable non-invasive method which should allow following genetic testing. For enrichment of foetal cells from peripheral maternal blood we combined Ficoll-Paque density gradient 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 using HLA (DRB1, DQB1, DQA1) and Polymarker System (LDLR, GYPA, HBGG, D7S8, GC) as genetic markers. Different paternal alleles of studied 8 loci were detected in 13 out of 19 samples of cells enriched from maternal peripheral blood between the 13th and 36th week of gestation. Our results demonstrate 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.
Keywords
nucleated red blood cells, CD71, maternal blood, high-gradient magnetic cell separation, MACS, paternal allele, HLA, polymarker system.
Funding
This work was supported by the Internal Czech Grant Agency, Ministry of Health, Czech Republic, project No. 4537-3, and by 2nd Medical Faculty, Charles University in Prague, project No. VZ 111300003.
References
Copyright
This is an open-access article distributed under the terms of the Creative Commons Attribution License.