Single-Color Digital PCR Provides High-Performance Detection of Cancer Mutations from Circulating DNA.

ABSTRACT: We describe a single-color digital PCR assay that detects and quantifies cancer mutations directly from circulating DNA collected from the plasma of cancer patients. This approach relies on a double-stranded DNA intercalator dye and paired allele-specific DNA primer sets to determine an absolute count of both the mutation and wild-type-bearing DNA molecules present in the sample. The cell-free DNA assay uses an input of 1 ng of nonamplified DNA, approximately 300 genome equivalents, and has a molecular limit of detection of three mutation DNA genome-equivalent molecules per assay reaction. When using more genome equivalents as input, we demonstrated a sensitivity of 0.10% for detecting the BRAF V600E and KRAS G12D mutations. We developed several mutation assays specific to the cancer drive

SUBMITTER: Wood-Bouwens C 

PROVIDER: S-EPMC6593258 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature