Rapid and Specific Screening Assay for KRAS Oncogene Mutation by a Novel Gene Amplification Method

Epidermal growth factor receptor (EGFR) is a target of molecular therapeutics for colorectal cancer. However, mutation of Kirsten rat sarcoma viral oncogene homolog (KRAS) gene at codons 12 and 13 attenuates the therapeutic effect of anti-EGFR therapies. Therefore, the detection of KRAS gene mutation is important for therapeutic decision-making.

KRAS gene mutation at codons 12 (c.34G>T, c.35G>C, c.35G>A) and 13 (c.38G>A) in six cancer cell lines were investigated. A loop-mediated isothermal amplification-based procedure was developed that employed peptide nucleic acid to suppress amplification of the wild-type allele.

This mutation-oriented gene-amplification procedure can amplify the DNA fragment of the KRAS gene with codon 12 and codon 13 mutation within 30 min. Moreover, boiled cells can work as template resources.

This newly developed procedure can be useful for patient stratification for anti-EGFR therapies.