title: Ultrasensitive single-nucleotide polymorphism detection using target-recycled ligation, strand displacement and enzymatic amplification.
abstract: We report herein the development of a highly sensitive and selective approach for label-free DNA detection by combining target-recycled ligation (TRL), magnetic nanoparticle assisted target capture/separation, and efficient enzymatic amplification. We show that our approach can detect as little as 30 amol (600 fM in 50 μL) of unlabelled single-stranded DNA targets and offer an exquisitely high discrimination ratio (up to >380 fold with background correction) between a perfect-match cancer mutant and its single-base mismatch (wild-type) DNA target. Furthermore, it can quantitate the rare cancer mutant (KRAS codon 12) in a large excess of coexisting wild-type DNAs down to 0.75%. This sensor appears to be well-suited for sensitive SNP detection and a wide range of DNA mutation based diagnostic applications.
authors: School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors: Yue
authors: Y
authors: Zhang
authors: Yuan
authors: Y
authors: Guo
authors: Philip
authors: P
authors: Quirke
authors: Dejian
authors: D
authors: Zhou
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D011088:DNA Ligases / Q000378:metabolism; D015342:DNA Probes / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000737:chemistry*; D005787:Gene Frequency; D006801:Humans; D058185:Magnetite Nanoparticles / Q000737:chemistry; D044367:Phase Transition; D020641:Polymorphism, Single Nucleotide; D044366:Transition Temperature
chemical_list: D015342:DNA Probes; D004277:DNA, Single-Stranded; D058185:Magnetite Nanoparticles; D004247:DNA; D011088:DNA Ligases
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