251. Systematic biases in reference-based plasma cell-free DNA fragmentomic profiling.
- Author
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Liu X, Yang M, Hu D, An Y, Wang W, Lin H, Pan Y, Ju J, and Sun K
- Subjects
- Humans, Neoplasms genetics, Neoplasms blood, Neoplasms diagnosis, Sequence Analysis, DNA methods, Liquid Biopsy methods, Bias, High-Throughput Nucleotide Sequencing methods, Cell-Free Nucleic Acids genetics, Cell-Free Nucleic Acids blood
- Abstract
Plasma cell-free DNA (cfDNA) fragmentation patterns are emerging directions in cancer liquid biopsy with high translational significance. Conventionally, the cfDNA sequencing reads are aligned to a reference genome to extract their fragmentomic features. In this study, through cfDNA fragmentomics profiling using different reference genomes on the same datasets in parallel, we report systematic biases in such conventional reference-based approaches. The biases in cfDNA fragmentomic features vary among races in a sample-dependent manner and therefore might adversely affect the performances of cancer diagnosis assays across multiple clinical centers. In addition, to circumvent the analytical biases, we develop Freefly, a reference-free approach for cfDNA fragmentomics profiling. Freefly runs ∼60-fold faster than the conventional reference-based approach while generating highly consistent results. Moreover, cfDNA fragmentomic features reported by Freefly can be directly used for cancer diagnosis. Hence, Freefly possesses translational merit toward the rapid and unbiased measurement of cfDNA fragmentomics., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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