cfDNA UniFlow: A unified preprocessing pipeline for cell-free DNA data from liquid biopsies

Abstract

AbstractBackgroundCell-free DNA (cfDNA), a broadly applicable biomarker commonly sourced from urine or blood, is extensively used for research and diagnostic applications. In various settings, genetic and epigenetic information is derived from cfDNA. However, a unified framework for its processing is lacking, limiting the universal application of innovative analysis strategies and the joining of data sets.FindingsHere, we describe cfDNA UniFlow, a unified, standardized, and ready-to-use workflow for processing cfDNA samples. The workflow is written in Snakemake and can be scaled from stand -alone computers to cluster environments. It includes methods for processing raw genome sequencing data as well as specialized approaches for correcting sequencing errors, filtering, and quality control. Sophisticated methods for detecting copy number alterations and estimating and correcting GC-related biases are readily incorporated. Furthermore, it includes methods for extracting, normalizing and visualizing coverage signals around user defined regions in case-control settings. Ultimately, all results and metrics are aggregated in a unified report, enabling easy access to a wide variety of information for further research and downstream analysis.ConclusionsWe provide an automated pipeline for processing cell-free DNA sampled from liquid biopsies, including a wide variety of additional functionalities like bias correction and signal extraction. With our focus on scalability and extensibility, we provide a foundation for future cfDNA research and faster clinical applications.Issue SectionTechnical NoteAvailability and implementationSource code and extensive documentation is available on our GitHub repository (https://github.com/kircherlab/cfDNA-UniFlow).