Data from: Analytical and clinical validation of a digital sequencing panel for quantitative, highly accurate evaluation of cell-free circulating tumor DNA

Next-generation sequencing of cell-free circulating solid tumor DNA
addresses two challenges in contemporary cancer care. First this method of
massively parallel and deep sequencing enables assessment of a
comprehensive panel of genomic targets from a single sample, and second,
it obviates the need for repeat invasive tissue biopsies. Digital
SequencingTM is a novel method for high-quality sequencing of circulating
tumor DNA simultaneously across a comprehensive panel of over 50
cancer-related genes with a simple blood test. Here we report the analytic
and clinical validation of the gene panel. Analytic sensitivity down to
0.1% mutant allele fraction is demonstrated via serial dilution studies of
known samples. Near-perfect analytic specificity (> 99.9999%)
enables complete coverage of many genes without the false positives
typically seen with traditional sequencing assays at mutant allele
frequencies or fractions below 5%. We compared digital sequencing of
plasma-derived cell-free DNA to tissue-based sequencing on 165 consecutive
matched samples from five outside centers in patients with stage III-IV
solid tumor cancers. Clinical sensitivity of plasma-derived NGS was 85.0%,
comparable to 80.7% sensitivity for tissue. The assay success rate on
1,000 consecutive samples in clinical practice was 99.8%. Digital
sequencing of plasma-derived DNA is indicated in advanced cancer patients
to prevent repeated invasive biopsies when the initial biopsy is
inadequate, unobtainable for genomic testing, or uninformative, or when
the patient’s cancer has progressed despite treatment. Its clinical
utility is derived from reduction in the costs, complications and delays
associated with invasive tissue biopsies for genomic testing.