Redefining "Gold Standard": Ultra-Sensitive Characterization Of Commercial Dna Standards With Duplex Sequencing

DNA standards are a critical resource in diagnostic and research labs. Positive and negative controls are essential for validating the sensitivity and specificity of next generation sequencing (NGS) and other genetic assays. Several commercial DNA standards have been widely used to validate clinical oncology assays. However, as genomic technologies have become more sensitive, the metrics defining what a “gold standard” entails have not been carefully re-evaluated. Cell culture and synthetic chemical means of generating DNA standards have the potential to artificially introduce mutations. Cell lines are often exposed to supraphysiologic levels of reactive oxygen. Oligonucleotide synthesis has an error rate orders of magnitude above biological levels. This background may not interfere with detection of clonal or sub clonal variants with moderately sensitive assays, but it will obscure the presence of very rare mutations when assessing with extremely sensitive ones and it will decrease apparent assay performance.