title: A novel single fluorophore-labeled double-stranded oligonucleotide probe for fluorescence-enhanced nucleic acid detection based on the inherent quenching ability of deoxyguanosine bases and competitive strand-displacement reaction.
abstract: We develop a novel single fluorophore-labeled double-stranded oligonucleotide (OND) probe for rapid, nanostructure-free, fluorescence-enhanced nucleic acid detection for the first time. We further demonstrate such probe is able to well discriminate single-base mutation in nucleic acid. The design takes advantage of an inherent quenching ability of guanine bases. The short strand of the probe is designed with an end-labeled fluorophore that is placed adjacent to two guanines as the quencher located on the long opposite strand, resulting in great quenching of dye fluorescence. In the presence of a target complementary to the long strand of the probe, a competitive strand-displacement reaction occurs and the long strand forms a more stable duplex with the target, resulting in the two strands of the probe being separated from each other. As a consequence of this displacement, the fluorophore and the quencher are no longer in close proximity and dye fluorescence increases, signaling the presence of target.
authors.affiliation: State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, Jilin, China.
authors.forename: Yingwei
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jingqi
authors.initials: J
authors.lastname: Tian
authors.forename: Hailong
authors.initials: H
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Xuping
authors.initials: X
authors.lastname: Sun
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D003849:Deoxyguanosine / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D008958:Models, Molecular; D009154:Mutation; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes / Q000737:chemistry* / Q000235:genetics; D012313:RNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D013050:Spectrometry, Fluorescence; D013997:Time Factors
chemical_list: D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D012313:RNA; D004247:DNA; D003849:Deoxyguanosine