title: Highly specific and sensitive electrochemical genotyping via gap ligation reaction and surface hybridization detection.
abstract: This paper developed a novel electrochemical genotyping strategy based on gap ligation reaction with surface hybridization detection. This strategy utilized homogeneous enzymatic reactions to generate molecular beacon-structured allele-specific products that could be cooperatively annealed to capture probes stably immobilized on the surface via disulfide anchors, thus allowing ultrasensitive surface hybridization detection of the allele-specific products through redox tags in close proximity to the electrode. Such a unique biphasic architecture provided a universal methodology for incorporating enzymatic discrimination reactions in electrochemical genotyping with desirable reproducibility, high efficiency and no interferences from interficial steric hindrance. The developed technique was demonstrated to show intrinsic high sensitivity for direct genomic analysis, and excellent specificity with discriminativity of single nucleotide variations.
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, PR China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.forename: Yan-Li
authors.initials: YL
authors.lastname: Zhang
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Xu
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Jiang
authors.forename: Guo-Li
authors.initials: GL
authors.lastname: Shen
authors.forename: Ru-Qin
authors.initials: RQ
authors.lastname: Yu
mesh_terms: D000483:Alleles; D001609:Beta-Globulins / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000235:genetics; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D005838:Genotype; D006046:Gold / Q000737:chemistry; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010084:Oxidation-Reduction; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity
chemical_list: D001609:Beta-Globulins; D006046:Gold; D004247:DNA