Detection of Genetic Variants in Human Leukocyte Antigen Genes

1 . A computer-implemented method of identifying somatic variants in a human leukocyte antigen (HLA) gene in a subject, the method comprising:

for an HLA gene,

obtaining a patient specific HLA reference genome from the HLA type data;

receiving HLA sequence read data from one or more normal samples and receiving HLA sequence read data for a tumor sample associated with a patient;

aligning the HLA sequence read data for the one or more normal samples and the HLA sequence data for the tumor sample to the patient specific HLA reference genome;

adjusting a mapping quality score associated with each read having a low mapping quality score indicative of multiple-mapping;

performing a variant calling process on the filtered aligned read data and determining the somatic variants of the HLA class; and

generating and storing a report of somatic variants of the HLA gene annotated for functional effect.

2 . The method of claim 1 , further comprising:

before determining the somatic variants of the HLA class, removing variants resulting from the variant calling process that are low confidence variants.

3 . The method of claim 2 , further comprising removing the variants by:

identifying, from a list of candidate somatic variants, variants in which all sequencing reads that support the variant map to the right of the variant or to the left of the variant.

4 . The method of claim 3 , further comprising; if the all the sequence reads map to the right or to the left of the variant, determining if the sequencing reads map to a different HLA gene and filtering the sequencing reads that map to the different HLA gene.

5 . The method of claim 1 , further comprising:

before determining the somatic variants of the HLA class, removing germline variants from variants identified by the variant calling process.

6 . The method of claim 5 , further comprising removing the germline variants comprises:

removing the germline variants from a list of candidate somatic variants, in response to tumor sample reads being below a threshold value.

7 . The method of claim 5 , further comprising removing the germline variants comprises:

removing the germline variants from a list of candidate somatic variants, in response to normal sample reads being above a threshold value.

8 . The method of claim 5 , further comprising removing the germline variants comprises:

removing the germline variants from a list of candidate somatic variants, in response to tumor variant allele frequency (VAF) and normal VAF being equal.

9 . The method of claim 5 , further comprising removing the germline variants comprises:

removing the germline variants from a list of candidate somatic variants, in response to a ratio of tumor variant allele frequency (VAF) to normal VAF being lower than a threshold value.

10 . The method of claim 1 , further comprising:

before determining the somatic variants of the HLA class, removing variants likely resulting from sequencing artifacts from variants identified by the variant calling process.

11 . The method of claim 11 , further comprising removing variants likely resulting from sequencing artifacts from variants identified by the variant calling process by:

removing variants from a list of candidate somatic variants, if all sequencing reads that support the variant map only to the forward strand or only to the reverse strand.

12 . The method of claim 1 , further comprising designating a variant as a somatic variant, if the ratio of tumor variant allele frequency (VAF) to normal VAF is greater than a threshold value.

13 . The method of claim 1 , wherein filtering the aligned read data to remove low quality reads by applying the event based filtering rule to the aligned read data comprises:

determining a mismatch event according to: mismatch event=edit distance−the size of insertions−the size of deletions;

determining an event count according: event count=mismatch event+number of insertions+number of deletions; and

outputting from the event based filtering rule the read data with the event count greater than 1 and removing all read data without the event counter greater than 1.

14 . The method of claim 1 , wherein the HLA class is HLA Class 1, and the somatic variants are located in one of the following genes: HLA-A, HLA-B, and HLA-C.

15 . The method of claim 1 , further comprising obtaining HLA type data that includes HLA reference sequence data corresponding to a plurality of different HLA alleles.

16 . The method of claim 1 , further comprising: updating the patient specific HLA reference genome to match the HLA sequence read data of the one or more normal samples.

17 . The method of claim 1 , wherein the one or more normal samples are associated with the patient.

18 . The method of claim 1 , wherein the one or more normal samples are selected from normal samples of non-patients determined to have a matching an HLA genotype of the patient.

19 . The method of claim 1 , further comprising obtaining the patient specific HLA reference genome by applying the HLA type data to a HLA typing module and generating, using the HLA typing module, the patient specific HLA reference genome.

20 . The method of claim 1 , further comprising, prior to performing the variant calling process on the filtered aligned read data, removing non-paired reads from the filtered aligned read data.

21 . The method of claim 20 , wherein removing non-paired reads from the filtered aligned read data comprises removing read data having an unmapped read end.

22 . The method of claim 1 , wherein adjusting the mapping quality score associated with each remaining read having the low mapping quality score indicative of multiple-mapping comprises setting an equal weight for each remaining read having the low mapping quality score indicative of multiple-mapping.