Rapid Detection of Clinically Important SNPs and Mutations by Two-color SmartAmp Method
Alexander Lezhava
Omics Sciences Center (OSC), RIKEN
Abstract:
Rapid, accurate and cost-effective patients tests for mutations and genetic polymorphism is a key requirement for the personalized therapy. We have recently developed single nucleotide polymorphism (SNP) detection method, named Smart Amplification Process (SmartAmp) that can successfully solve afore-mentioned requirements. SmartAmp is able detect SNPs or mutations in 30 to 45min under isothermal conditions directly from whole blood without any DNA purification step.
Furthermore, we introduced a new DNA labeling and detection method, the so-called Exciton Dye Primers, which have specific advantages for real-time monitoring of isothermal amplification methods. Dyes quenched by exciton coupling principle are attached to the single nucleotide of the primer and emit flurescence only after hybridization to the complementary sequence. Testing in SmartAmp assay, our Exciton Dye Primers showed very high signal specificity as compared to SYBR GreenI, a commonly used intercalating dye for real-time monitoring. Introduction of new labeling using different dyes, allowed us to proceeding multiplex reaction in one tube, saving labor time and reagent cost. We are showing further specific examples for the use of our new two-color method for genotyping SNPs relevant to Warfarin dosing and EGFR mutation detection.
