Diacarta Ltd

Aiguo Zhang

Michael J Powell

Paul Okunieff

Lurong Zhang

Michael Y Sha

Lulu Zhang

Ke Zhan

Elena Peletskaya

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.

DNA mutation detection employing enrichment of mutant polynucleotide sequences and minimally invasive sampling

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.

This invention provides methods and systems for detecting sequence variants in a sample nucleic acid. Methods include PNA clamping/PCR using primers with binding moieties followed by capture and detection of amplicons on a solid support. Systems include PCR reagents with primers having binding moieties, a PNA clamping probe, a solid support capable of capturing the PCR amplicons incorporating the binding moieties, and a detector device.

Detection of PNA clamping

This invention relates generally to methods for detecting cell damage as a consequence of pathophysiological or traumatic insults such as in a nuclear accident, bioterror attack, tumorigenesis, infections or in individuals with cardiovascular disease.

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San Ramon, CA, United States of America

Aiguo Zhang