Uracil-DNA glycosylase-engineered recombinase polymerase amplification for single-nucleotide polymorphism discrimination and genotyping.

Abstract

We developed an isothermal, equipment-minimal single-nucleotide polymorphism genotyping platform coupling recombinase polymerase amplification with toehold-mediated strand displacement. By using dU-primers and uracil-DNA glycosylase treatment to expose toeholds on amplicons, we achieved 0.1% mutant allele sensitivity and full concordance with standard genotyping methods across all tested clinical samples, providing a robust solution for potential application.