DNA mutation detection employing enrichment of mutant polynucleotide sequences and minimally invasive sampling
US11208689B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 12, 2019 |
| Grant date | Dec 28, 2021 |
| Priority date | — |
| Expiry date | Jul 12, 2039 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2531/113
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The invention relates to a method for enriching a target polynucleotide sequence containing a genetic variation said method comprising: (a) providing two primers targeted to said target polynucleotide sequence; (b) providing a target specific xenonucleic acid clamp oligomer specific for a wildtype polynucleotide sequence; (c) generating multiple amplicons using PCR under specific temperature cycling conditions; and (d) detecting said amplicons. We introduce a novel molecule, Xenonucleic Acid (XNA) for the NGS library preparation. XNA is able to selectively suppress amplification of DNA with wild type alleles and amplify DNA containing mutant alleles. Mutants with low allelic frequency will be easily detectable without deep sequencing after enrichment by adding XNA in multiplex PCR. The 17 actionable mutants related to lung or colorectal cancer diseases at different variant allelic frequency (VAF) % were investigated. Clinical sensitivity is significantly improved with XNA in various types of samples.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.