MAESTRO affords ‘breadth and depth’ for mutation testing

Abstract

The ability to assay large numbers of low-abundance mutations is crucial in biomedicine. Yet, the technical hurdles of sequencing multiple mutations at extremely high depth and accuracy remain daunting. For sequencing low-level mutations, it’s either ‘depth or breadth’ but not both. Here, we report a simple and powerful approach to accurately track thousands of distinct mutations with minimal reads. Our technique called MAESTRO (minor allele enriched sequencing through recognition oligonucleotides) employs massively-parallel mutation enrichment to empower duplex sequencing—one of the most accurate methods—to track up to 10,000 low-frequency mutations with up to 100-fold less sequencing. In example use cases, we show that MAESTRO could enable mutation validation from cancer genome sequencing studies. We also show that it could track thousands of mutations from a patient’s tumor in cell-free DNA, which may improve detection of minimal residual disease from liquid biopsies. In all, MAESTRO improves the breadth, depth, accuracy, and efficiency of mutation testing.

Cell-free DNA technologies for the analysis of brain cancer

“…Although tumour-guided sequencing is not suitable for diagnostic and screening purposes [44,48], its potential to detect relapse and pseudoprogression in glioma remains unexplored. Tumour-guided sequencing could be further combined with mutational enrichment methods (via size selection or thermodynamic-based strategies), which even if they have not been tested in the glioma context, could have a strong potential for improving the monitoring of relapse in these patients [49,50].…”