Journal article

Analytical validation of an error-corrected ultra-sensitive ctDNA next-generation sequencing assay

Heidi Fettke, Jason A Steen, Edmond M Kwan, Patricia Bukczynska, Shivakumar Keerthikumar, David Goode, Maria Docanto, Nicole Ng, Luciano Martelotto, Christine Hauser, Melissa C Southey, Arun A Azad, Tu Nguyen-Dumont

BioTechniques: the journal of laboratory technology for bioresearch | FUTURE SCI LTD | Published : 2020

Abstract

Plasma circulating tumor DNA (ctDNA) analysis has emerged as a minimally invasive means to perform molecular tumor typing. Here we developed a custom ultra-sensitive ctDNA next-generation sequencing assay using molecular barcoding technology and off-the-shelf reagents combined with bioinformatics tools for enhanced ctDNA analysis. Assay performance was assessed via a spike-in experiment and the technique was applied to analyze 41 plasma samples from men with advanced prostate cancer. Orthogonal validation was performed using a commercial assay. Sensitivity and specificity of 93 and 99.5% were recorded for ultra-rare somatic variants (<1%), with high concordance observed between the in-house ..

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