Multiplexed Assays for Detection of Mutations in PIK3CA

phoinositide-3-kinase, catalytic, alpha polypeptide) have recently been described in a number of cancers, and their detection is currently limited because of the low sensitivity of conventional sequencing techniques. METHODS: We combined Amplification Refractory Mutation System (ARMS™; AstraZeneca) allele-spe-cific PCR and Scorpions ™ (DxS) to develop assays for tumor-borne PIK3CA mutations and used real-time PCR to develop high-throughput multiplexed assays for the most commonly reported PIK3CA mutants (H1047L, H1047R, E542K, E545K). RESULTS: These assays were more sensitive than se-quencing and could detect 5 copies of mutant DNA in proportions as low as 0.1 % of the total DNA. We as-sayed DNA extracted from human tumors and de