Analysis of Base-Position Error Rate of Next-Generation Sequencing to Detect Tumor Mutations in Circulating DNA

摘要: BACKGROUND: Detecting single-nucleotide variations and insertions/deletions in circulating tumor DNA is challenging because of their low allele frequency. The clinical use to characterize genetic alterations requires new methods based on next-generation sequencing. METHODS: We developed a method quantification error rate each base position [position (PER)]. To identify mutations, binomial test was used compare the minor-allele frequency measured PER at position. This process validated control samples 373 plasma from patients with lung or pancreatic cancer. RESULTS: Minimal mutated frequencies were 0.003 for 0.001 insertions/deletions. Independent testing performed by droplet digital PCR (n = 231 samples) showed strong agreement base-PER (κ 0.90). CONCLUSIONS: Targeted sequencing analyzed represents robust cost detect