Clinical validation of KRAS, BRAF, and EGFR mutation detection using next-generation sequencing.

ABSTRACT:

Objectives

To validate next-generation sequencing (NGS) technology for clinical diagnosis and to determine appropriate read depth.

Methods

We validated the KRAS, BRAF, and EGFR genes within the Ion AmpliSeq Cancer Hotspot Panel using the Ion Torrent Personal Genome Machine (Life Technologies, Carlsbad, CA).

Results

We developed a statistical model to determine the read depth needed for a given percent tumor cellularity and number of functional genomes. Bottlenecking can result from too few input genomes. By using 16 formalin-fixed, paraffin-embedded (FFPE) cancer-free specimens and 118 cancer specimens with known mutation status, we validated the six traditional analytic performance characteristics recommended by the Next-Generation Sequencing: Standardization of Clin

SUBMITTER: Lin MT 

PROVIDER: S-EPMC4332779 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature