Detection of low prevalence somatic mutations in solid tumors with ultra-deep targeted sequencing

Genome Biol. 2011 Dec 20;12(12):R124. doi: 10.1186/gb-2011-12-12-r124.


Ultra-deep targeted sequencing (UDT-Seq) can identify subclonal somatic mutations in tumor samples. Early assays' limited breadth and depth restrict their clinical utility. Here, we target 71 kb of mutational hotspots in 42 cancer genes. We present novel methods enhancing both laboratory workflow and mutation detection. We evaluate UDT-Seq true sensitivity and specificity (> 94% and > 99%, respectively) for low prevalence mutations in a mixing experiment and demonstrate its utility using six tumor samples. With an improved performance when run on the Illumina Miseq, the UDT-Seq assay is well suited for clinical applications to guide therapy and study clonal selection in heterogeneous samples.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Animals
  • Automation, Laboratory
  • Carcinoma / diagnosis*
  • Carcinoma / genetics
  • Databases, Genetic
  • Genes, Neoplasm / genetics*
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Mice
  • Middle Aged
  • Mutation Rate
  • Mutation*
  • Sarcoma / diagnosis*
  • Sarcoma / genetics
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods*
  • Xenograft Model Antitumor Assays