DNA was extracted from paraffin blocks of 126 CRC patients. KRAS codon 12/13 and BRAF V600E mutational status was assessed using high resolution melting (HRM), direct sequencing (DS) of the HRM polymerase chain reaction (PCR) product. In addition, the Therascreen Amplification Refractory Mutation System (ARMS)-Scorpion KRAS assay and BRAF pyrosequencing were employed; both assays claim to require less tumour cells in comparison with DS.
KRAS and BRAF were found to be mutually exclusive. Mutation frequencies were 33.9%for KRAS, and for BRAF 19.0%, respectively. Concordance of KRAS mutational status between biopsy and resection specimens was 97.4%(ARMS), 98.4%(DS) and 99.2%(HRM), respectively. For BRAF concordance was 98.4%(Pyro, DS) and 99.2%(HRM).
KRAS and BRAF mutational status of endoscopic biopsies and resection specimens of CRC showed a >95%concordance. Endoscopic biopsies can be confidently used for molecular analysis.