Bisulfite-converted duplexes for the strand-specific detection and quantification of rare mutations

Study ID Alternative Stable ID Type
EGAS00001002406 Other

Study Description

The identification of mutations that are present at low frequencies in clinical samples is an essential component of precision medicine. The development of molecular barcoding for next generation sequencing has greatly enhanced the sensitivity of detecting such mutations by massively parallel sequencing. However, further improvements in specificity would be useful for a variety of applications. We herein describe a technology (BiSeqS) that can increase the specificity of sequencing by at least two orders of magnitude over and above that achieved with molecular barcoding and can be applied to any massively parallel sequencing instrument. BiSeqS employs bisulfite treatment to distinguish the two strands of molecularly barcoded DNA; its specificity arises from the requirement for the same mutation to be identified in both strands. Because no library preparation is required, the technology permits very efficient use of the template DNA as well as sequence reads, which are nearly all confined to the amplicons of interest. Such efficiency is critical for clinical samples, such as plasma, in which only tiny amounts of DNA are often available. We show here that BiSeqS can be applied to evaluate transversions, as well as small insertions or deletions, and can reliably detect one mutation among >10,000 wild type molecules.

Study Datasets 1 dataset.

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Dataset ID Description Technology Samples
EGAD00001003323 Runs that contain data for the sensitivity and specificity experiments for BiSeqS. Illumina MiSeq 2

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Publications Citations
Bisulfite-converted duplexes for the strand-specific detection and quantification of rare mutations.
Proc Natl Acad Sci U S A 114:2017 4733-4738