Personalized methods for detecting circulating tumor DNA
The present disclosure relates to a laboratory execution system that provides for automation of laboratory processes. A centralized data management system may be dynamically updated and used to facilitate management of components of the laboratory execution system, such as an automation system and an analytics results management system that may facilitate complex analytical functions, such as synthesizing raw test data. Potential workflows include the detection of specific molecules of interest.
1. A personalized method for detecting circulating tumor DNA in a patient comprising:
(a) obtaining genomic DNA from solid tumor tissue from a patient and sequencing the genomic DNA from the solid tumor tissue;
(b) preparing a set of oligonucleotides comprising a signature panel of mutations, said signature panel being specific for said patient and comprising a set of greater than 50 tumor-specific somatic mutations, wherein (i) said mutations in said signature panel of mutations comprise one or more mutations selected from SNPs, insertions, deletions, and translocations, (ii) wherein tumor-specific somatic mutations are obtained by aligning the genomic DNA from the tumor to a reference human genome that is not from the patient thereby obtaining a plurality of tumor-specific mutations, and (iii) wherein the tumor-specific somatic mutations are present in genomic DNA from the tumor but not present in genomic DNA from a non-tumor tissue from the patient; and
(c) contacting cell free DNA (cfDNA) obtained from a fluid sample from the patient with the set of oligonucleotides comprising said signature panel to enrich a fraction of the cfDNA corresponding to the signature panel, and sequencing the fraction of cfDNA to detect circulating tumor DNA (ctDNA) in the fluid sample, wherein ctDNA is determined to be present when one or more sequence reads corresponding to one or more of the tumor-specific somatic mutations in the signature panel is detected.
2. The method of claim 1 , wherein the non-tumor tissue from the patient is matched tissue to the tumor.
3. The method of claim 1 , wherein sequencing the genomic DNA from the solid tumor tissue comprises whole genome sequencing or targeted sequencing.
4. The method of claim 1 , further repeating (c) at one or more times either:
(i) during treatment to determine the efficacy of said treatment, or
(ii) following completion of treatment to determine recurrence of cancer.
5. The method of claim 1 , wherein said fluid sample comprises blood plasma.
6. The method of claim 3 , wherein targeted sequencing comprises sequencing exons.
7. A personalized method for detecting circulating tumor DNA in a patient comprising:
(a) obtaining genomic DNA from solid tumor tissue from a patient and sequencing the genomic DNA from the solid tumor tissue;
(b) preparing a set of oligonucleotides comprising a signature panel of mutations, said signature panel being specific for said patient and comprising a set of greater than 50 tumor-specific somatic mutations, wherein (i) said mutations in said signature panel of mutations comprise one or more mutations selected from SNPs, insertions, deletions, and translocations, (ii) wherein tumor-specific mutations are obtained by aligning the genomic DNA from the tumor to a reference human genome that is not from the patient, and (iii) wherein the tumor-specific somatic mutations are present in genomic DNA from the tumor but not present in genomic DNA from a non-tumor tissue from the patient; and
(c) contacting cell free DNA (cfDNA) obtained from a fluid sample from the patient with the set of oligonucleotides comprising said signature panel to enrich a fraction of the cfDNA corresponding to the signature panel, and sequencing the fraction of cfDNA, to detect circulating tumor DNA (ctDNA) in the fluid sample, wherein ctDNA is detected by the presence of one or more sequence reads corresponding to one or more of the tumor-specific somatic mutations in the signature panel.
8. The method of claim 7 , wherein the non-tumor tissue from the patient is matched tissue to the tumor.
9. The method of claim 7 , wherein and sequencing the genomic DNA from the solid tumor tissue comprises whole genome sequencing or targeted sequencing.
10. The method of claim 7 , further repeating (c) at one or more times either:
(i) during treatment to determine the efficacy of said treatment, or
(ii) following completion of treatment to determine recurrence of cancer.
11. The method of claim 7 , wherein said fluid sample comprises blood plasma.
12. The method of claim 9 , wherein targeted sequencing comprises sequencing exons.
13. A personalized method for detecting circulating tumor DNA in a patient comprising:
(a) obtaining genomic DNA from solid tumor tissue from a patient and sequencing the genomic DNA from the solid tumor tissue;
(b) preparing a set of oligonucleotides comprising a signature panel of mutations, said signature panel being specific for said patient and comprising a set of greater than 50 tumor-specific somatic mutations, wherein (i) said mutations in said signature panel of mutations comprise one or more mutations selected from SNPs, insertions, deletions, and translocations (ii) wherein tumor-specific mutations are obtained by aligning the genomic DNA from the tumor to a reference human genome that is not from the patient, and (iii) wherein the tumor-specific somatic mutations are present in genomic DNA from the tumor but not present in genomic DNA from a non-tumor tissue from the patient; and
(c) contacting cell free DNA (cfDNA) obtained from a fluid sample from the patient with the set of oligonucleotides comprising said signature panel to enrich a fraction of the cfDNA corresponding to the signature panel, and sequencing the fraction of cfDNA, thereby detecting circulating tumor DNA (ctDNA) in the fluid sample when one or more sequence reads corresponding to one or more of the tumor-specific somatic mutations in the signature panel is detected.
14. The method of claim 13 , wherein the non-tumor tissue from the patient is matched tissue to the tumor.
15. The method of claim 13 , wherein and sequencing the genomic DNA from the solid tumor tissue comprises whole genome sequencing or targeted sequencing.
16. The method of claim 13 , further repeating (c) at one or more times either:
(i) during treatment to determine the efficacy of said treatment, or
(ii) following completion of treatment to determine recurrence of cancer.
17. The method of claim 13 , wherein said fluid sample comprises blood plasma.
18. The method of claim 15 , wherein targeted sequencing comprises sequencing exons.