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Electrochemical DNA detection via exonuclease and target-catalyzed transformation of surface-bound probes.

ABSTRACT: We report a single-step, single-reagent, label-free, isothermal electrochemical DNA sensor based on the phenomenon of target recycling. The sensor exploits strand-specific exonuclease activity to achieve the selective enzymatic digestion of target/probe duplexes. This results in a permanent change in the probe structure that yields an increased faradaic current and liberates the intact target molecule to interact with additional detection probes to achieve further signal amplification. Using this architecture, we achieve an improved detection limit in comparison to hybridization-based sensors without amplification. We also demonstrate a 16-fold signal amplification factor at low target concentrations. Combined with the advantages of electrochemical detection and its ready integration with microelectronics, our approach may represent a promising path toward direct DNA detection at the point of care.

SUBMITTER: Hsieh K

PROVIDER: S-EPMC2883016 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature

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Electrochemical DNA detection via exonuclease and target-catalyzed transformation of surface-bound probes.

Hsieh Kuangwen K Xiao Yi Y Tom Soh H H

Langmuir : the ACS journal of surfaces and colloids 20100601 12

We report a single-step, single-reagent, label-free, isothermal electrochemical DNA sensor based on the phenomenon of target recycling. The sensor exploits strand-specific exonuclease activity to achieve the selective enzymatic digestion of target/probe duplexes. This results in a permanent change in the probe structure that yields an increased faradaic current and liberates the intact target molecule to interact with additional detection probes to achieve further signal amplification. Using thi ...[more]

PMID: 20232926

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Project description:In the last decade, researchers have been searching for innovative platforms, methods, and techniques able to address recurring problems with the current cancer detection methods. Early disease detection, fast results, point-of-care sensing, and cost are among the most prevalent issues that need further exploration in this field. Herein, studies are focused on overcoming these problems by developing an electrochemical device able to detect telomerase as a cancer biomarker. Electrochemical platforms and techniques are more appealing for cancer detection, offering lower costs than the established cancer detection methods, high sensitivity inherent to the technique, rapid signal processing, and their capacity of being miniaturized. Therefore, Au interdigital electrodes and electrochemical impedance spectroscopy were used to detect telomerase activity in acute T cell leukemia. Different cancer cell concentrations were evaluated, and a detection limit of 1.9 × 105 cells/mL was obtained. X-ray photoelectron spectroscopy was used to characterize the telomerase substrate (TS) DNA probe self-assembled monolayer on gold electrode surfaces. Atomic force microscopy displayed three-dimensional images of the surface to establish a height difference of 9.0 nm between the bare electrode and TS-modified Au electrodes. The TS probe is rich in guanines, thus forming secondary structures known as G-quadruplex that can be triggered with a fluorescence probe. Confocal microscopy fluorescence images showed the formation of DNA G-quadruplex because of TS elongation by telomerase on the Au electrode surface. Moreover, electrodes exposed to telomerase containing 2',3'-dideoxyguanosine-5'-triphosphate (ddGTP) did not exhibit high fluorescence, as ddGTP is a telomerase inhibitor, thus making this device suitable for telomerase inhibitors capacity studies. The electrochemical method and Au microchip device may be developed as a biosensor for a point-of-care medical device.

Dataset Information

Electrochemical DNA detection via exonuclease and target-catalyzed transformation of surface-bound probes.

Publications

Electrochemical DNA detection via exonuclease and target-catalyzed transformation of surface-bound probes.

Similar Datasets

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