Highly specific and sensitive electrochemical genotyping via gap ligation reaction and surface hybridization detection.
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.
Huang
Yong
Y
State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, PR China.
Zhang
Yan-Li
YL
Xu
Xiangmin
X
Jiang
Jian-Hui
JH
Shen
Guo-Li
GL
Yu
Ru-Qin
RQ
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
D001609:Beta-Globulins; D006046:Gold; D004247:DNA