ccfDNA In the Lab: Optimizing Purification for Sequencing
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Margaret L. Gulley MD is professor and Director of Molecular Pathology in the Department of Pathology and Laboratory Medicine at University of North Carolina at Chapel Hill, USA. She provides laboratory services to patients with cancer, heritable disease, infectious disease, or transplant. Dr. Gulley’s research on virus-related malignancy seeks to validate novel molecular assays that add value for disease classification and monitoring. Work with professional societies aims to promote high quality molecular services in clinical laboratories.
Tuesday, September 10, 2019
ccfDNA Webinar Series: The Basics and Beyond
Circulating cell-free DNA (ccfDNA) can be used to monitor tumor burden and monitor disease progress.
In Part 3 of 3 ccfDNA webinars, Dr. Gulley discusses variables important to isolating high-quality ccfDNA from plasma samples including anticoagulant, plasma volume and storage length and conditions.
Register for the previous parts in this series:
Part 1 - ccfDNA 101: Emerging Trends in Oncology Research
Part 2 - ccfDNA Workflows: Honing in on the Target
Additional Webinar Information:
Circulating cell-free DNA assays serve to monitor tumor burden and to track clonal evolution, including emerging drug resistance clones. There are many important pre-analytic variables for assuring high quality sequencing results on plasma DNA, including choice of anticoagulant, input plasma volume, extraction chemistry, and the duration and storage conditions of blood before blood is processed to separate plasma. Interfering substances or processes may include hemolysis, leukocyte lysis, and nucleases. Assay validation resources can help in assay design and experimental design to optimize sensitivity, specificity, linearity and reproducibility of sequencing results. Controls, calibrators, and quality checks are valuable tools to demonstrate assays perform as expected. The adverse impact of tumor heterogeneity on tissue-based genomic profiles may be overcome, in part, by testing plasma that may yield a more global or systemic portrait of tumor clones and emerging subclones. Monitoring patients in serial plasma tests performed over time may be more informative than test results at a single timepoint. Ongoing research continues to reveal the utility of plasma DNA as a tool to enhance early diagnosis, tumor classification, and patient response to therapeutic interventions.