This work was supported by the Danish Blood Donors' Research Foundation, the Danish National Research Foundation, and the Lundbeck Foundation.
NEW METHODS FOR BLOOD GROUP GENOTYPING
Next-generation sequencing: proof of concept for antenatal prediction of the fetal Kell blood group phenotype from cell-free fetal DNA in maternal plasma
Article first published online: 3 APR 2013
© 2013 Department of Clinical Immunology, University Hospital Copenhagen. Transfusion © 2013 American Association of Blood Banks
Special Issue: Twenty Years since the Cloning of the Blood Group Genes
Volume 53, Issue 11pt2, pages 2892–2898, November 2013
How to Cite
Rieneck, K., Bak, M., Jønson, L., Clausen, F. B., Krog, G. R., Tommerup, N., Nielsen, L. K., Hedegaard, M. and Dziegiel, M. H. (2013), Next-generation sequencing: proof of concept for antenatal prediction of the fetal Kell blood group phenotype from cell-free fetal DNA in maternal plasma. Transfusion, 53: 2892–2898. doi: 10.1111/trf.12172
- Issue published online: 12 NOV 2013
- Article first published online: 3 APR 2013
- Manuscript Accepted: 23 JAN 2013
- Manuscript Revised: 2 JAN 2013
- Manuscript Received: 25 OCT 2012
- Danish Blood Donors' Research Foundation
- Danish National Research Foundation
- Lundbeck Foundation
Maternal immunization against KEL1 of the Kell blood group system can have serious adverse consequences for the fetus as well as the newborn baby. Therefore, it is important to determine the phenotype of the fetus to predict whether it is at risk. We present data that show the feasibility of predicting the fetal KEL1 phenotype using next-generation sequencing (NGS) technology.
Study Design and Methods
The KEL1/2 single-nucleotide polymorphism was polymerase chain reaction (PCR) amplified with one adjoining base, and the PCR product was sequenced using a genome analyzer (GAIIx, Illumina); several millions of PCR sequences were analyzed.
The results demonstrated the feasibility of diagnosing the fetal KEL1 or KEL2 blood group from cell-free DNA purified from maternal plasma.
This method requires only one primer pair, and the large amount of sequence information obtained allows well for statistical analysis of the data. This general approach can be integrated into current laboratory practice and has numerous applications. Besides DNA-based predictions of blood group phenotypes, platelet phenotypes, or sickle cell anemia, and the determination of zygosity, various conditions of chimerism could also be examined using this approach. To our knowledge, this is the first report focused on antenatal blood group determination using NGS.