Multiplexed assays for detection of mutations in PIK3CA.

摘要: Background: Mutations in the PIK3CA gene (phosphoinositide-3-kinase, catalytic, alpha polypeptide) have recently been described a number of cancers, and their detection is currently limited because low sensitivity conventional sequencing techniques. Methods: We combined Amplification Refractory Mutation System (ARMS™; AstraZeneca) allele-specific PCR Scorpions™ (DxS) to develop assays for tumor-borne mutations used real-time high-throughput multiplexed most commonly reported mutants (H1047L, H1047R, E542K, E545K). Results: These were more sensitive than could detect 5 copies mutant DNA proportions as 0.1% total DNA. assayed extracted from human tumors detected mutation frequencies 10.2% colorectal cancer, 38.7% breast 1.9% lung 2.9% melanoma. In contrast, only 53% by our assay. Conclusions: Multiplexed assays, which can easily be applied clinical samples, developed mutations.