%0 Journal Article %A Jonathan C. M. Wan %A Katrin Heider %A Davina Gale %A Suzanne Murphy %A Eyal Fisher %A James Morris %A Florent Mouliere %A Dineika Chandrananda %A Andrea Marshall %A Andrew B. Gill %A Pui Ying Chan %A Emily Barker %A Gemma Young %A Wendy N. Cooper %A Irena Hudecova %A Francesco Marass %A Graham R. Bignell %A Constantine Alifrangis %A Mark R. Middleton %A Ferdia A. Gallagher %A Christine Parkinson %A Amer Durrani %A Ultan McDermott %A Christopher G. Smith %A Charles Massie %A Pippa G. Corrie %A Nitzan Rosenfeld %T High-sensitivity monitoring of ctDNA by patient-specific sequencing panels and integration of variant reads %D 2019 %R 10.1101/759399 %J bioRxiv %P 759399 %X Circulating tumor-derived DNA (ctDNA) can be used to monitor cancer dynamics noninvasively. Patients with small tumors have few copies of ctDNA in plasma, resulting in limited sensitivity to detect low-volume or residual disease. We show that sampling limitations can be overcome and sensitivity for ctDNA detection can be improved by massively parallel sequencing when hundreds to thousands of mutations are identified by tumor genotyping. We describe the INtegration of VAriant Reads (INVAR) analysis pipeline, which combines patient-specific mutation lists with both custom error-suppression methods and signal enrichment based on biological features of ctDNA. In this framework, the sensitivity can be estimated independently for each sample based on the number of informative reads, which is the product of the number of mutations analyzed and the average depth of unique sequencing reads. We applied INVAR to deep sequencing data generated by custom hybrid-capture panels, and showed that when ~106 informative reads were obtained INVAR allowed detection of tumor-derived DNA fractions to parts per million (ppm). In serial samples from patients with advanced melanoma on treatment, we detected ctDNA when imaging confirmed tumor volume of ~1cm3. In patients with resected early-stage melanoma, ctDNA was detected in 40% of patients who later relapsed, with higher rates of detection when more informative reads were obtained. We further demonstrated that INVAR can be generalized and allows improved detection of ctDNA from whole-exome and low-depth whole-genome sequencing data. %U https://www.biorxiv.org/content/biorxiv/early/2019/09/11/759399.full.pdf