Comparative analysis of various targeted enrichment methodologies to identify actionable mutations in lung cancer.

e13110Background: The recent expansion of knowledge about genetic changes associated with various diseases including cancers, using whole genome or exome sequencing has led to the development of focused or targeted gene panels. Such panels can be used to characterize both germ-line and somatic changes with a high degree of specificity and sensitivity. Two different approaches-hybridization capture and PCR-have been widely used to generate target specific libraries covering relevant loci. The limited amount of sample available, particularly in tissue biopsies and FFPE specimens, can restrict the use of both approaches for investigating a large number of gene targets. In addition, false positives may occur during the early steps of PCR or during whole genome amplification introducing potential errors. Methods: We used probes specific for the coding regions of 215 genes from Nimblegen (capture approach), NuGEN (AMP approach) and Illumina (PCR approach) to identify clinically actionable mutations and gene fus...