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Fetal RHD Genotyping in Maternal Plasma at 11–13 Weeks of Gestation

Akolekar R.a · Finning K.b · Kuppusamy R.a · Daniels G.b · Nicolaides K.H.a, c
aHarris Birthright Research Centre for Fetal Medicine, King’s College Hospital, London, bBristol Institute for Transfusion Sciences and International Blood Group Reference Laboratory, NHS Blood and Transplant, Bristol, and cDepartment of Fetal Medicine, University College Hospital, London, UK Fetal Diagn Ther 2011;29:301–306 (DOI:10.1159/000322959)


Objective: To examine the feasibility of fetal RHD genotyping at 11–13 weeks’ gestation from analysis of circulating cell-free fetal DNA (ccffDNA) in the plasma of RhD negative pregnant women using a high-throughput robotic technique. Methods: Stored plasma (0.5 ml) from 591 RhD negative women was used for extraction of ccffDNA by a robotic technique. Real-time quantitative polymerase chain reaction (PCR) with probes for exons 5 and 7 of the RHD gene was then used to determine the fetal RHD genotype, which was compared to the neonatal RhD phenotype. Results: In total there were 502 (85.7%) cases with a conclusive result and 84 (14.3%) with an inconclusive result. The prenatal test predicted that the fetus was RhD positive in 332 cases and in all of these the prediction was correct, giving a positive predictive value of 100% (95% CI 96.8–100). The test predicted that the fetus was RhD negative in 170 cases and in 164 of these the prediction was correct, giving a negative predictive value for RhD positive fetuses of 96.5% (95% CI 93.7–99.2). Conclusion: The findings demonstrate the feasibility and accuracy of non-invasive fetal RHD genotyping at 11–13 weeks with a high-throughput technique.


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