Digital Sequencing Using Liquid Biopsies: Clinical and Technical Aspects

Digital Sequencing Using Liquid Biopsies: Clinical and Technical Aspects

Abstract
The clinical need to detect and quantify mutant allele variants at low frequency in challenging sample types, such as liquid biopsies, is increasing. Analysis of circulating nucleic acids requires optimized and standardized workflows, including sampling, preanalytics, nucleic acids analysis, data analysis and clinical interpretation. Digital sequencing of nucleic acids using unique molecular identifiers enables close to error-free sequencing and minimal amplification biases. Here, we will present a standardized workflow for handling blood samples for nucleic acids analysis, including the development of different quality controls. We will also demonstrate how digital sequencing can be used to analyze both DNA and RNA using different sample types. Robust digital sequencing requires well-performing assays and we will demonstrate the importance of high quality primers. Finally, we will show data from several clinical studies related to melanomas and sarcomas and how analysis of blood-based biomarkers can assist in cancer management.