title: Evaluation of Nine Somatic Variant Callers for Detection of Somatic Mutations in Exome and Targeted Deep Sequencing Data.
abstract: Next generation sequencing is extensively applied to catalogue somatic mutations in cancer, in research settings and increasingly in clinical settings for molecular diagnostics, guiding therapy decisions. Somatic variant callers perform paired comparisons of sequencing data from cancer tissue and matched normal tissue in order to detect somatic mutations. The advent of many new somatic variant callers creates a need for comparison and validation of the tools, as no de facto standard for detection of somatic mutations exists and only limited comparisons have been reported. We have performed a comprehensive evaluation using exome sequencing and targeted deep sequencing data of paired tumor-normal samples from five breast cancer patients to evaluate the performance of nine publicly available somatic variant callers: EBCall, Mutect, Seurat, Shimmer, Indelocator, Somatic Sniper, Strelka, VarScan 2 and Virmid for the detection of single nucleotide mutations and small deletions and insertions. We report a large variation in the number of calls from the nine somatic variant callers on the same sequencing data and highly variable agreement. Sequencing depth had markedly diverse impact on individual callers, as for some callers, increased sequencing depth highly improved sensitivity. For SNV calling, we report EBCall, Mutect, Virmid and Strelka to be the most reliable somatic variant callers for both exome sequencing and targeted deep sequencing. For indel calling, EBCall is superior due to high sensitivity and robustness to changes in sequencing depths.
authors.affiliation: Department of Clinical Genetics, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense, Denmark.|Human Genetics, Institute of Clinical Research, University of Southern Denmark, Winsløvparken 19, 5000, Odense, Denmark.
authors.forename: Anne Bruun
authors.initials: AB
authors.lastname: Krøigård
authors.affiliation: Department of Clinical Genetics, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense, Denmark.|Human Genetics, Institute of Clinical Research, University of Southern Denmark, Winsløvparken 19, 5000, Odense, Denmark.
authors.forename: Mads
authors.initials: M
authors.lastname: Thomassen
authors.affiliation: Department of Pathology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark.
authors.forename: Anne-Vibeke
authors.initials: AV
authors.lastname: Lænkholm
authors.affiliation: Department of Clinical Genetics, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense, Denmark.|Human Genetics, Institute of Clinical Research, University of Southern Denmark, Winsløvparken 19, 5000, Odense, Denmark.
authors.forename: Torben A
authors.initials: TA
authors.lastname: Kruse
authors.affiliation: Department of Clinical Genetics, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense, Denmark.|Human Genetics, Institute of Clinical Research, University of Southern Denmark, Winsløvparken 19, 5000, Odense, Denmark.
authors.forename: Martin Jakob
authors.initials: MJ
authors.lastname: Larsen
mesh_terms: D000465:Algorithms; D001483:Base Sequence; D018270:Carcinoma, Ductal, Breast / Q000235:genetics*; D019295:Computational Biology / Q000379:methods*; D059472:Exome / Q000235:genetics*; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D009154:Mutation / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*