Monitoring mutations using prior knowledge of variants

A method comprising:

• (a) obtaining, by a data processing system, sequence data for a plurality of target regions in a sample of cell free DNA from a subject, wherein the plurality of target regions are selected from a plurality of genomic regions that comprise a plurality of known variants;
• (b) querying, by the data processing system, the sequence data for one or more variants of the plurality of known variants;
• (c) calculating, by the data processing system, a first number of mutant molecules for each of the queried one or more variants based on an allele fraction for each of the queried one or more variants;
• (d) modeling, by the data processing system, background in the sample of cell free DNA, wherein the modeling comprises randomly sampling the sequence data a predetermined number of times for one or more types of variants present in the plurality of genomic regions and calculating a second number of mutant molecules for each of the one or more variants in the random samples based on an allele fraction for each of the one or more variants in the random samples;
• (e) comparing, by the data processing system, the second number of mutant molecules for each of the one or more variants in the random samples to the first number of mutant molecules for each of the queried one or more variants;
• (f) generating, by the data processing system, a ratio based on the comparison, wherein the ratio is:
• (a number of the random samples where the second number of mutant molecules is greater than the first number of mutant molecules):(the predetermined number of times), and the ratio is a probability value for a null-hypothesis that the second number of mutant molecules of the background is greater than the first number of mutant molecules of the queried variant;
• (g) determining, by the data processing system, a variable significance value based on a number of the queried one or more variants while maintaining a predetermined false discovery rate, wherein the determining the variable significance value comprises:
• (i) determining a different significance value for each number of the one or more variants of the plurality of known variants that are capable of being queried while maintaining the predetermined false discovery rate, and selecting the significance value for the number of the queried one or more variants from the determined different significance values;
• or (ii) determining an equation relating the significance value to the number of the queried one or more variants while maintaining a predetermined false discovery rate;
• (h) determining, by the data processing system, a tumor burden for the subject based on the first number of mutant molecules for each of the queried one or more variants;
• and
• (i) determining, by the data processing system, a statistical significance of the tumor burden based on the probability value and the variable significance value.

A method of diagnosing a patient with minimal residual disease comprising:

• (a) detecting, by a data processing system, one or more variants in a sample of cell free DNA from a subject, wherein the one or more variants are selected from a plurality of variants known to be specific to a tumor or disease area of the subject;
• (b) counting, by the data processing system, the detected one or more variants;
• (c) determining, by the data processing system, a tumor burden based on the count of the one or more variants;
• (d) determining, by the data processing system, a statistical significance of the tumor burden, wherein the determining the statistical significance comprises:
• (i) modeling background noise using a multinucleotide context of base changes from randomly selected positions in sequence data for the sample of cell free DNA to define an empirically derived probability value, (ii) determining a variable significance value based on a number of the detected one or more variants while maintaining a predetermined false discovery rate, and (iii) comparing the probability value to the variable significance value, where the lower the probability value compared to the variable significance value, the higher the statistical significance of the tumor burden;
• and
• (e) determining, by the data processing system, whether the subject has minimal residual disease based on the statistical significance of the tumor burden.

A method of performing a survivability analysis, the method comprising:

• (a) detecting, by a data processing system, one or more variants in a sample of cell free DNA from a subject, wherein the one or more variants are selected from a plurality of variants known to be specific to a tumor or disease area of the subject;
• (b) counting, by the data processing system, the detected one or more variants;
• (c) determining, by the data processing system, a statistical significance of the count of the one or more variants, wherein the determining the statistical significance comprises:
• (i) modeling background noise using a multinucleotide context of base changes from randomly selected positions in sequence data for the sample of cell free DNA to define an empirically derived probability value, (ii) determining a variable significance value based on a number of the detected one or more variants while maintaining a predetermined false discovery rate, and (iii) comparing the probability value to the variable significance value, where the lower the probability value compared to the variable significance value, the higher the statistical significance of the count of the one or more variants;
• (d) repeating steps (a)-(c) for a predetermined number of samples of cell free DNA from the subject at various points prior to and during a treatment regimen to obtain the count of the one or more variants for each of the samples of cell free DNA and the statistical significance associated with each of the counts of the one or more variants;
• and
• (e) predicting, by the data processing system, a clinical outcome of the treatment regimen for the subject based upon the count of the one or more variants for each of the samples of cell free DNA and the statistical significance associated with each of the counts of the one or more variants.