Electrocatalytic nucleic acid hybridization detection
First Claim
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1. A method of detecting nucleic acid hybridization between a nucleic acid probe and a target nucleic acid in a sample, the method comprising:
- (a) providing a nucleic acid probe immobilized on nanoelectrode ensembles;
(b) contacting under hybridization conditions the nanoelectrode ensembles and the immobilized nucleic acid probe with the sample and a redox pair comprising (i) a nucleic acid binding compound comprising a first transition metal complex that interacts electrostatically with the nucleic acid probe, and (ii) a redox-active probe comprising a second transition metal complex;
wherein the sample and redox pair are in solution; and
(c) measuring an electrocatalytic signal generated by hybridization of the nucleic acid probe and the target nucleic acid in the sample, wherein an increase of the signal detected relative to a signal of a control sample comprising no target nucleic acid, indicates hybridization between the nucleic acid probe and the target nucleic acid, and wherein a decrease in the signal detected relative to that of perfect complementarity between the nucleic acid probe and the target nucleic acid indicates a mismatch between the nucleic acid probe and the target nucleic acid.
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Abstract
The detection of specific nucleic acid sequences using electrochemical readout would permit the rapid and inexpensive detection and identification of bacterial pathogens and the analysis of human genes. A new assay developed for this purpose is described that harnesses an electrocatalytic process to monitor nucleic acid hybridization. Furthermore, the new assay when used on nanoscale electrodes, provides ultrasensitive detection of nucleic acids.
48 Citations
32 Claims
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1. A method of detecting nucleic acid hybridization between a nucleic acid probe and a target nucleic acid in a sample, the method comprising:
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(a) providing a nucleic acid probe immobilized on nanoelectrode ensembles; (b) contacting under hybridization conditions the nanoelectrode ensembles and the immobilized nucleic acid probe with the sample and a redox pair comprising (i) a nucleic acid binding compound comprising a first transition metal complex that interacts electrostatically with the nucleic acid probe, and (ii) a redox-active probe comprising a second transition metal complex;
wherein the sample and redox pair are in solution; and(c) measuring an electrocatalytic signal generated by hybridization of the nucleic acid probe and the target nucleic acid in the sample, wherein an increase of the signal detected relative to a signal of a control sample comprising no target nucleic acid, indicates hybridization between the nucleic acid probe and the target nucleic acid, and wherein a decrease in the signal detected relative to that of perfect complementarity between the nucleic acid probe and the target nucleic acid indicates a mismatch between the nucleic acid probe and the target nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method of detecting nucleic acid hybridization between a first nucleic acid and a second nucleic acid, the method comprising:
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(a) providing the first nucleic acid immobilized on nanoelectrode ensembles; (b) contacting under hybridization conditions the nanoelectrode ensembles and the immobilized first nucleic acid with a solution suspected of containing the second nucleic acid and containing a redox pair comprising (i) a first transition metal complex that interacts electrostatically with the nucleic acids, and (ii) a redox-active probe comprising a second transition metal complex; and (c) measuring an electrocatalytic signal generated by hybridization of the first nucleic acid and the second nucleic acid, wherein an increase of the signal detected relative to a signal of a control sample comprising no second nucleic acid, indicates that the nucleic acid hybridization between the first nucleic acid and the second nucleic acid, and wherein a decrease in the signal detected relative to that of perfect complementarity between the first and second nucleic acids indicates a mismatch between the first nucleic acid and the second nucleic acid. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18)
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19. A method of detecting nucleic acid hybridization between a nucleic acid probe and a target nucleic acid in a sample, the method comprising:
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(a) providing a nucleic acid probe immobilized on nanoelectrode ensembles; (b) contacting under hybridization conditions the nanoelectrode ensembles and the immobilized nucleic acid probe with the sample and a redox pair comprising a first transition metal complex that interacts electrostatically with the nucleic acid probe, and a redox-active probe comprising an ascorbic acid or tripropylamine, wherein the sample and the redox pair are in solution; and (c) measuring an electrocatalytic signal generated by hybridization of the nucleic acid probe and the target nucleic acid in the sample, wherein an increase of the signal detected relative to a signal of a control sample comprising no target nucleic acid, indicates that the nucleic acid hybridization between the nucleic acid probe and the target nucleic acid, and wherein a decrease in the signal detected relative to that of perfect complementarity between the nucleic acid probe and the target nucleic acid indicates a mismatch between the nucleic acid probe and the target nucleic acid. - View Dependent Claims (20, 21, 22, 23, 24, 25)
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26. A method of detecting nucleic acid hybridization between a first nucleic acid and a second nucleic acid, the method comprising:
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(a) providing the first nucleic acid immobilized on nanoelectrode ensembles; (b) contacting under hybridization conditions the nanoelectrode ensembles and the immobilized first nucleic acid with a solution suspected of containing the second nucleic acid and containing a redox pair comprising (i) a first transition metal complex that interacts electrostatically with the nucleic acids, and (ii) a redox-active probe comprising an ascorbic acid or tripropylamine; and (c) measuring an electrocatalytic signal generated by hybridization of the first nucleic acid and the second nucleic acid, wherein an increase of the signal detected relative to a signal of a control sample comprising no second nucleic acid, indicates that the nucleic acid hybridization between the first nucleic acid and the second nucleic acid, and wherein a decrease in the signal detected relative to that of perfect complementarity between the first and second nucleic acids indicates a mismatch between the first nucleic acid and the second nucleic acid. - View Dependent Claims (27, 28, 29, 30, 31, 32)
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Specification