Detecting Rare Mutations in Cell-Free DNA by Integrating an Ultrafast Amplicon-Based Targeted Library Preparation Method with Double-Stranded Unique Molecular Identifiers

摘要

The use of as cell-free DNA (cfDNA) for testing has seen tremendous growth in recent years, as liquid biopsy is a noninvasive and easily obtainable sample type with many diagnostics and prognostic values. Liquid biopsy can enable new applications, but it also presents challenges toward accurate variant detection due to the low fraction of mutant DNA present in theses samples, which can be easily buried by the artifacts and background noise from systematic PCR and sequencing errors, leading to false negative results. While some progress has been made by ligating adapters with unique molecular identifiers (UMIs) in hybrid capture-based methods followed by deep sequencing, such approaches have time-consuming, complicated, and tedious workflows, often resulting in disappointingly low on-target rates. Additionally, the large amount of cfDNA required for hybrid capture workflows is not realistic to obtain from patients.