Enhanced Sensitivity of Early Fetal Genetics using the QIAamp Circulating Nucleic Acid Kit

Multiple gestation represents the principle complication with in vitro fertilization (IVF), causing increased maternal and neonatal morbidity compared with singleton pregnancies, and estimated to cost $1 billion annually in the U.S. Better methods of embryo selection are needed before routine single embryo transfer protocols can be successful. Studies involving the evaluation of the predictive value of new embryo selection methodologies require the ability to track which of the multiple IVF embryos transferred actually implanted. DNA fingerprinting has therefore become an extremely powerful tool in the development of markers of embryonic reproductive potential. We previously demonstrated that cell-free fetal DNA enriched from maternal circulation can be used to determine which embryo implanted at 9 weeks gestation, avoiding invasive methodologies such as CVS or amniocentesis, and providing for more rapid clinical trials (Treff et al. 2011, Molecular Human Reproduction, 17, 434–438). As a surrogate for performance of DNA enrichment, analysis of fetal gender using quantitative real time PCR for 9 Y chromosome loci was more recently performed here after using either the QIAamp Circulating Nucleic Acid Kit or QIAamp Virus Kit. Maternal peripheral blood at 9 weeks gestation from 40 patients was studied and compared to genders observed upon delivery. Assay specific sensitivity to the presence of Y chromosome DNA was significantly better using the Circulating Nucleic Acid Kit compared to the QIAamp Virus Kit (95% compared to 83%, respectively). Specificity for samples from pregnancies with a male fetus was 100% for DNA enriched using either kit. In conclusion, the QIAamp Circulating Nucleic Acid Kit provides enhanced sensitivity to cell-free fetal DNA from maternal circulation and provides an opportunity for increasing the quantity and quality of downstream applications.

Nathan Treff

Dr. Nathan Treff serves as Director of Molecular Biology Research at Reproductive Medicine Associates of New Jersey, Assistant Professor at the Robert Wood Johnson Medical School, and Adjunct Professor at Rutgers University. Dr. Treff’s laboratory uses molecular genetics to discover, develop, and translate novel molecular diagnostics to improve the quality of life for patients with infertility. His research was awarded the 2007 and 2010 General Program Prizes and the 2009 and 2010 Society for Assisted Reproductive Technology Prize by the American Society for Reproductive Medicine. He completed his Ph.D. in biochemistry at Washington State University where he earned the Charles Glenn King Scholarship and the National Institutes of Health Predoctoral Fellowship in protein biotechnology. Dr. Treff trained as a postdoctoral fellow in stem cell biology at the University of Wisconsin-Madison and in reproductive biology at the EMD-Serono Research Institute.