Methods and systems for genetic analysis

Patent metadata

Field	Value
Publication number	US-12371746-B2
Application number	US-202418824319-A
Country	US
Kind code	B2
Filing date	Sep 4, 2024
Priority date	Jan 17, 2013
Publication date	Jul 29, 2025
Grant date	Jul 29, 2025

Abstract

This disclosure provides systems and methods for sample processing and data analysis. Sample processing may include nucleic acid sample processing and subsequent sequencing. Some or all of a nucleic acid sample may be sequenced to provide sequence information, which may be stored or otherwise maintained in an electronic storage location. The sequence information may be analyzed with the aid of a computer processor, and the analyzed sequence information may be stored in an electronic storage location that may include a pool or collection of sequence information and analyzed sequence information generated from the nucleic acid sample. Methods and systems of the present disclosure can be used, for example, for the analysis of a nucleic acid sample, for producing one or more libraries, and for producing biomedical reports. Methods and systems of the disclosure can aid in the diagnosis, monitoring, treatment, and prevention of one or more diseases and conditions.

First claim

What is claimed is:

1. A method of analyzing nucleic acid samples obtained from an individual comprising:
(a) generating a first subset of nucleic acid molecules from a first nucleic acid sample obtained from the individual by contacting the first nucleic acid sample with one or more pulldown probe sets that selectively enrich for an exome, wherein at least one of the one or more pulldown probe sets comprise 10 or more pulldown probes with different sequences;
(b) conducting a first sequencing assay on the first subset of nucleic acid molecules to yield a first result comprising a first nucleic acid sequence;
(c) producing, with the aid of a computer processor, one or more primer sets, wherein the one or more primer sets are configured to selectively amplify a plurality of polymorphisms, wherein the plurality of polymorphisms are based on the first result obtained in (b);
(d) generating a second subset of nucleic acid molecules from a second nucleic acid sample of the individual by selectively amplifying nucleic acid molecules with the one or more primer sets produced in step (c) to produce a set of amplicons, wherein the selectively amplifying comprises multiplex PCR; and
(e) conducting a second sequencing assay on the set of amplicons to yield a second result comprising a second nucleic acid sequence, thereby analyzing the nucleic acid samples.

2. The method of claim 1 , further comprising, subsequent to (e), generating a biomedical report that includes biomedical information of the subject, which biomedical information is indicative of the analysis.

3. The method of claim 2 , wherein the biomedical information of the subject is predictive, prognostic, or diagnostic of one or more biomedical features selected from the group consisting of: disease state, genetic risk of disease, efficacy of a drug therapy, and prediction of optimal drug dosage.

4. The method of claim 3 , wherein the disease state comprises cancer.

5. The method of claim 4 , wherein the cancer is a recurrent and/or refractory cancer.

6. The method of claim 1 , wherein individual instances of the pulldown probes comprised in the one or more pulldown probe sets each comprise between about 10 to about 500 nucleotides.

7. The method of claim 1 , wherein individual instances of the pulldown probes comprised in the one or more pulldown probe sets further comprise a label, wherein the label comprises biotin or a magnetic particle.

8. The method of claim 1 , wherein individual instances of the primers comprised in the one or more primer sets further comprise a sample identifier, wherein the sample identifier comprises a barcode.

9. The method of claim 1 , wherein the exome comprises: (i) exons, (ii) untranslated regions (UTRs), (iii) one or more splice sites, (iv) one or more intronic regions, and (v) one or more regulatory regions.

10. The method of claim 1 , wherein the first nucleic acid sample and/or the second nucleic acid sample comprises DNA, RNA, DNA/RNA hybrids, or cDNA derived from RNA.

11. The method of claim 1 , wherein the first result yielded by the first sequencing assay produces a set of nucleic acid sequencing information comprising a set of variant calls, wherein the set of variant calls comprises single nucleotide polymorphisms (SNPs) with heterozygous allelic forms.

12. The method of claim 1 , wherein the plurality of polymorphisms comprises: (i) known biomedically interpretable variants associated with a disease or indication, or (ii) one or more insertions, deletions, single nucleotide mutations, or a combination thereof.

13. The method of claim 12 , wherein the known biomedically interpretable variants associated with a disease or indication are selected from the group consisting of: (i) p53 mutations, (ii) Rb mutations, (iii) cell cycle regulators, (iv) cell cycle receptors, (v) cell cycle kinases, (vi) genes associated with cancer, and (vii) a combination thereof.

14. The method of claim 1 , wherein the first sequencing assay produces at least 10,000,000 sequencing reads.

15. The method of claim 1 , wherein the first sequencing assay generates at least 5,000,000 sequencing reads per run.

16. The method of claim 1 , wherein mean sequencing size of the first subset of nucleic acid molecules and/or the second subset of nucleic acid molecules is at least about 50 bases or base pairs to about 300 bases or base pairs.

17. The method of claim 1 , wherein the first sequencing assay and/or the second sequencing assay generates at least about 30 base pairs to about 300 base pairs per read.

18. The method of claim 1 , wherein the first subset of nucleic acid molecules and the second subset of nucleic acid molecules are derived from two or more different nucleic acid samples.

19. The method of claim 18 , wherein the two or more different nucleic acid samples are collected over two or more time points.

20. The method of claim 1 , wherein the first subset of nucleic acid molecules is derived from a tissue biopsy.

21. The method of claim 1 , wherein the second subset of nucleic acid molecules is derived from a sample selected from the group consisting of blood, plasma, and a blood fraction.

22. The method of claim 1 , wherein the first subset of nucleic acid molecules and/or the second subset of nucleic acid molecules is derived from a sample selected from the group consisting of a tissue biopsy, blood, plasma, and a blood fraction.

23. The method of claim 1 , further comprising, repeating steps (d)-(e) on a subsequently obtained nucleic acid sample from the individual.

24. The method of claim 23 , wherein the subsequently obtained nucleic acid sample is derived from a sample selected from the group consisting of blood, plasma, and a blood fraction.

25. The method of claim 23 , wherein the subsequently obtained nucleic acid sample comprises DNA, RNA, DNA/RNA hybrids, or cDNA derived from RNA.

26. The method of claim 1 , wherein the generating of step (a) further comprises:
(A) hybridizing at least part of the first nucleic acid sample with the one or more pulldown probe sets; and
(B) separating pulldown probe-hybridized nucleic acid molecules from pulldown probe-free nucleic acid molecules.

27. The method of claim 26 , wherein the generating of step (a) further comprises:
(C) conducting one or more elution reactions on the pulldown probe-hybridized nucleic acid molecules.

28. The method of claim 1 , wherein the first sequencing assay comprises sequencing by synthesis.

29. The method of claim 1 , wherein the second sequencing assay comprises sequencing by synthesis.

30. The method of claim 1 , wherein percent error of the second sequencing assay is less than about 0.001%.

Assignees

• Personalis Inc

Inventors

• Bartha Gabor T
• Chandratillake Gemma
• Chen Richard
• Garcia Sarah
• Lam Hugo Yu Kor
• Pratt Mark R
• West John

Classifications

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• G16B35/10 Primary

  Design of libraries · CPC title

• G16B20/20

  Allele or variant detection, e.g. single nucleotide polymorphism [SNP] detection · CPC title

• G16B20/10

  Ploidy or copy number detection · CPC title

• G16C20/60

  In silico combinatorial chemistry · CPC title