Abstract
Purpose
To examine the potential high throughput capability and efficiency of an automated DNA extraction system in combination with mass spectrometry for the non-invasive determination of the foetal Rhesus D status.
Methods
A total of 178 maternal plasma samples from RHD-negative pregnant women were examined, from which DNA was extracted using the automated Roche MagNA Pure™ system. Presence of the foetal RHD gene was detected by PCR for RHD exon 7 and subsequent analysis using the Sequenom MassArray™ mass spectrometric system.
Results
We determined that as little as 15 pg of RHD-positive genomic DNA could be detected in a background of 585 pg of RHD-negative genomic DNA. The analysis of the clinical samples yielded a sensitivity and specificity of 96.1 and 96.1%, respectively.
Conclusion
Our study indicated that automated DNA extraction in combination with mass spectrometry permits the determination of foetal Rhesus D genotype with an accuracy comparable to the current approaches using real-time PCR.
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Acknowledgments
Funding from the European Commission for the Special Non-invasive Advances in Fetal and Neonatal Evaluation (SAFE) Network of Excellence (LSHB-CT-2004-503243) and the Swiss National Science Foundation (SNSF 3200B0-107697/1), from which this study was partially funded, is gratefully acknowledged.
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Grill, S., Banzola, I., Li, Y. et al. High throughput non-invasive determination of foetal Rhesus D status using automated extraction of cell-free foetal DNA in maternal plasma and mass spectrometry. Arch Gynecol Obstet 279, 533–537 (2009). https://doi.org/10.1007/s00404-008-0774-5
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DOI: https://doi.org/10.1007/s00404-008-0774-5