Systems and methods to detect rare mutations and copy number variation
First Claim
1. A method for detecting a presence or absence of one or more somatic genetic variants in a sample comprising cell-free deoxyribonucleic acid (cfDNA) molecules from a subject having a cancer or suspected of having a cancer, the method comprising:
- (a) contacting a set of molecular barcodes comprising 2 to 1,000 different molecular barcodes with a population of cfDNA molecules obtained from the sample to produce tagged parent polynucleotides,wherein the tagged parent polynucleotides have a first molecular barcode from the set of molecular barcodes that is attached to a first end of a cfDNA molecule from among the population of cfDNA molecules and a second molecular barcode from the set of molecular barcodes that is attached to a second end of the cfDNA molecule, andwherein a plurality of the tagged parent polynucleotides has identical first molecular barcodes and second molecular barcodes;
(b) amplifying a plurality of the tagged parent polynucleotides to produce amplified progeny polynucleotides;
(c) sequencing a plurality of the amplified progeny polynucleotides to produce a set of sequencing reads; and
(d) detecting, from among a plurality of the set of sequencing reads, the presence or absence of one or more somatic genetic variants comprising a single nucleotide variant (SNV), a copy number variation (CNV), an insertion or deletion (indel), or a gene fusion.
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Abstract
The present disclosure provides a system and method for the detection of rare mutations and copy number variations in cell free polynucleotides. Generally, the systems and methods comprise sample preparation, or the extraction and isolation of cell free polynucleotide sequences from a bodily fluid; subsequent sequencing of cell free polynucleotides by techniques known in the art; and application of bioinformatics tools to detect rare mutations and copy number variations as compared to a reference. The systems and methods also may contain a database or collection of different rare mutations or copy number variation profiles of different diseases, to be used as additional references in aiding detection of rare mutations, copy number variation profiling or general genetic profiling of a disease.
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Citations
30 Claims
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1. A method for detecting a presence or absence of one or more somatic genetic variants in a sample comprising cell-free deoxyribonucleic acid (cfDNA) molecules from a subject having a cancer or suspected of having a cancer, the method comprising:
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(a) contacting a set of molecular barcodes comprising 2 to 1,000 different molecular barcodes with a population of cfDNA molecules obtained from the sample to produce tagged parent polynucleotides, wherein the tagged parent polynucleotides have a first molecular barcode from the set of molecular barcodes that is attached to a first end of a cfDNA molecule from among the population of cfDNA molecules and a second molecular barcode from the set of molecular barcodes that is attached to a second end of the cfDNA molecule, and wherein a plurality of the tagged parent polynucleotides has identical first molecular barcodes and second molecular barcodes; (b) amplifying a plurality of the tagged parent polynucleotides to produce amplified progeny polynucleotides; (c) sequencing a plurality of the amplified progeny polynucleotides to produce a set of sequencing reads; and (d) detecting, from among a plurality of the set of sequencing reads, the presence or absence of one or more somatic genetic variants comprising a single nucleotide variant (SNV), a copy number variation (CNV), an insertion or deletion (indel), or a gene fusion. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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Specification