Accurate detection of KRAS NRAS and BRAF mutations in metastatic colorectal cancers by bridged nucleic acid-clamp real-time PCR

摘要

Experimental design. We tested FFPE samples from 50 patients with colorectal cancer. Tissue sections were subjected to PCR-rSSO and BNA-clamp PCR. BNA-clamp PCR involved real-time PCR and amplified the mutated allele. The amplification plot was verified using mutation-positive samples and an internal control was amplified to confirm assay integrity. PCR products from BNA-clamp PCR were subsequently analyzed by Sanger sequencing to confirm the nucleotide changes