Use of DNA thermodynamics for low-abundance mutation detection by DNA hybridization

Abstract

Mutation detection is of great importance in many biological processes and diagnostics. Single-nucleotide variations are common, where a single base-pair is substituted during replication of wild-type DNA. Duplex formation via hybridisation provides an important detection technique where one of the strands is designed to match the mutant sequence and probe its presence. The probe is, however, subject to cross-hybridisation of wild-type DNA, lowering detection sensitivity in case of low-abundant mutants in a majority wild-type background. We use thermodynamics-based probe design and Langmuir theory to increase sensitivity and quantify the amount of mutant in a sample.