The Use of Maternal Plasma for Prenatal RhD Blood Group Genotyping

  • Kirstin Finning
  • Pete Martin
  • Geoff Daniels
Part of the METHODS IN MOLECULAR BIOLOGY™ book series (MIMB, volume 496)


Alloimmunization to the blood group antibody anti-RhD (anti-D) is the most common cause of hemolytic disease of the fetus and newborn. Knowledge of fetal D type in women with anti-D makes management of the pregnancy much easier and avoids unnecessary procedures in those women with a D-negative fetus. Fetal D typing can be performed by detection of an RHD gene in cell-free DNA in the plasma of D-negative pregnant women. The technology involves real-time quantitative polymerase chain reactions targeting exons 4, 5, and 10 of RHD, with the exons 4 and 10 tests performed as a multiplex. Testing for SRY in multiplex with the RHD exon 5 test provides an internal control for the presence of fetal DNA when the fetus is male. Fetal D typing has become the standard of care in England in pregnant women with a significant level of anti-D.

Key Words

Blood groups Rh fetal testing free fetal DNA hemolytic disease of the fetus and newborn 


  1. 1.
    Klein, H. G., Anstee, D. J. (2005) Blood Transfusion in Clinical Medicine, 11th edition. Blackwell Publishing, Oxford, UK.Google Scholar
  2. 2.
    National Institute for Clinical Excellence (2002) Technology Appraisal Guidance 41. Guidance on the Use of Routine Antenatal Anti-D Prophylaxis for RhD-Negative Women. NICE, London, UK.Google Scholar
  3. 3.
    van der Schoot, C. E., Tax, G. H. M., Rijnders, R. J. P., de Haas, M., Christiaens, G. C. M. L. (2003) Prenatal typing of Rh and Kell blood group system antigens: the edge of a watershed. Transfus Med Rev 17, 31–44.CrossRefPubMedGoogle Scholar
  4. 4.
    Daniels, G., Finning, K., Martin, P., Soothill, P. (2004) Fetal blood group genotyping from DNA from maternal plasma: an important advance in the management and prevention of haemolytic disease of the fetus and newborn. Vox Sang 87, 225–232.CrossRefPubMedGoogle Scholar
  5. 5.
    Daniels, G., Finning, K., Martin, P., Summers, J. (2006) Fetal blood group genotyping. Present and future. Ann NY Acad Sci 1075, 88–95.CrossRefPubMedGoogle Scholar
  6. 6.
    Lo, Y. M., Tein, M. S. C., Lau, T. K., Haines, C. J., Leung, T. N., Poon, P. M. K., et al. (1998) Quantitative analysis of fetal DNA in maternal plasma and serum: implications for noninvasive prenatal diagnosis. Am J Hum Genet 62, 768–775.CrossRefPubMedGoogle Scholar
  7. 7.
    Finning, K. M., Martin, P. G., Soothill, P. W., Avent, N. D. (2002) Prediction of fetal D status from maternal plasma: introduction of a new noninvasive fetal RHD genotyping service. Transfusion 42, 1079–1085.CrossRefPubMedGoogle Scholar
  8. 8.
    Finning, K., Martin, P., Summers, J., Daniels, G. (2007) Fetal genotyping for the K (Kell) and Rh C, c, and E blood groups on cell-free fetal DNA from maternal plasma. Transfusion 47, 2126–2133.CrossRefPubMedGoogle Scholar
  9. 9.
    Daniels, G. (2002) Human Blood Groups, 2nd edition. Oxford: Blackwell Science.Google Scholar
  10. 10.
    Colin, Y., Chérif-Zahar, B., Le Van Kim, C., Raynal, V., Van Huffel, V., Cartron, J-P. (1991) Genetic basis of the RhD-positive and RhD-negative blood group polymorphism as determined by Southern analysis. Blood 78, 2747–2752.Google Scholar
  11. 11.
    Singleton, B. K., Green, C. A., Avent, N. D., Martin, P. G., Smart, E., Daka, A., et al. (2000) The presence of an RHD pseudogene containing a 37 base pair duplication and a nonsense mutation in most Africans with the Rh D-negative blood group phenotype. Blood 95, 12–18.PubMedGoogle Scholar
  12. 12.
    Faas, B. H. W., Beckers, E. A. M., Wildoer, P., Ligthart, P. C., Overbeeke, M. A., Zondervan, H. A., et al. (1997) Molecular background of VS and weak C expression in blacks. Transfusion 37, 38–44.CrossRefPubMedGoogle Scholar
  13. 13.
    Page-Christiaens, G. C., Bossers, B., van der Schoot, C. E., De Haas, M. (2006) Use of bi-allelic insertion/deletion polymorphisms as a positive control for fetal genotyping in maternal blood: first clinical experience. Ann NY Acad Sci 1075, 123–129.CrossRefPubMedGoogle Scholar
  14. 14.
    Chan, K. C., Ding, C., Gerovassili, A., Yeung, S. W., Chiu, R. W., Leung, T. N., et al. (2006) Hypermethylated RASSF1A in maternal plasma: A universal fetal DNA marker that improves the reliability of noninvasive prenatal diagnosis. Clin Chem 52, 2211–2218.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Kirstin Finning
    • 1
  • Pete Martin
    • 1
  • Geoff Daniels
    • 1
  1. 1.International Blood Group Reference LaboratoryNHS Blood and TransplantUK

Personalised recommendations