Extended RAS and BRAF Mutation Analysis Using Next-Generation Sequencing.

摘要: Somatic mutations in KRAS, NRAS, and BRAF genes are related to resistance anti-EGFR antibodies colorectal cancer. We have established an extended RAS mutation assay using a next-generation sequencer analyze these mutations. Multiplexed deep sequencing was performed detect somatic within BRAF, including minor mutated components. first validated the technical performance of multiplexed 10 normal DNA 20 formalin-fixed, paraffin-embedded (FFPE) tumor samples. To demonstrate potential clinical utility our assay, we profiled 100 FFPE samples 15 plasma obtained from cancer patients. used variant calling approach based on Poisson distribution. The distribution mutation-positive population hypothesized follow distribution, status defined as value greater than significance level error rate (α = 2 x 10-5). cut-off determined be average plus 7 standard deviations. Mutation analysis specimens without any invalid cases. Mutations were detected at frequency 59% (59/100). KRAS concordance between this Scorpion-ARMS 92% (92/100). also analyzed. identified both tissue 6 genetic screening for detection clinically relevant liquid