Abstract 1872: A novel method for efficient and hands-free purification of circulating DNA from human plasma

Abstract

Abstract Circulating or Cell-Free DNA in plasma can be used to detect biomarkers that show great promise for diagnosis and monitoring of cancer, and is already being used as a non-invasive method to detect trisomy in fetuses. There is currently great interest in the discovery of new cancer biomarkers and their potential clinical application. However, reproducible and efficient purification of these highly fragmented and low-concentration species represents a major challenge. Here we will present a novel method which is completely automated and allows parallel purification of circulating nucleic acid from plasma and serum using a medium-throughput robot. Sixteen samples can be processed simultaneously. The method was optimized to produce high-quality DNA that is suitable for use in quantitative PCR and next generation sequencing. In addition, the absence of any pre-processing steps improves reproducibility and lowers the risk of contamination.Initial characterization on plasma from pregnant women showed that fetal DNA could be detected as early as 4 weeks into gestation and could be tracked throughout pregnancy. Further characterization showed that the system was able to reliably detect less than 25 copies/ml plasma of fragmented DNA that was spiked into the sample. Subsequent work on plasma from patients with colorectal cancer showed that the system was able to detect DNA containing both wild-type and mutated EGFR, suggesting that this method can be a useful tool when screening plasma samples for biomarkers of interest. Citation Format: Douglas White, Douglas Horejsh, Zhiyang Zeng, Tetsuo Uyeda, Poncho Meisenheimer, Marjeta Urh. A novel method for efficient and hands-free purification of circulating DNA from human plasma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1872. doi:10.1158/1538-7445.AM2014-1872