Abstract

The general clinical procedure for viral DNA detection or gene mutation diagnosis following polymerase chain reaction (PCR) often involves gel electrophoresis and DNA sequencing, which is usually time-consuming. In this study, we have proposed a facile strategy to construct a DNA biosensor, in which the platinum electrode was modified with a dual-film of electrochemically synthesized poly(3,4-ethylenedioxythiophene) (PEDOT) resulting in immobilized gold nanoparticles, with the gold nanoparticles easily immobilized in a uniform distribution. The DNA probe labeled with a SH group was then assembled to the fabricated electrode and employed to capture the target DNA based on the complementary sequence. The hybridization efficiency was evaluated with differential pulse voltammetry (DPV) in the presence of daunorubicin hydrochloride. Our results demonstrated that the peak current in DPV exhibited a linear correlation the concentration of target DNA that was complementary to the probe DNA. Moreover, the electrode could be reused by heating denaturation and re-hybridization, which only brought slight signal decay. In addition, the addition of the oxidized form of nicotinamide adenine dinucleotide (NAD+) could dramatically enhance the sensitivity by more than 5.45-fold, and the limit-of-detection reached about 100 pM.

Highlights

  • Recent research in DNA biosensor fabrication has gained remarkable progress by offering reliable approaches to quantify DNA and detect mutations [1,2,3]

  • We have reported a PEDOT modified DNA biosensor that is capable of quantifying ssDNA, detecting mutations, and even the dsDNA product from a polymerase chain reaction (PCR) reaction

  • With the aim of achieving real-time monitoring of DNA coupled with a PCR reaction, the DNA biosensor is expected achieving real-time monitoring of DNA coupled with a PCR reaction, the DNA biosensor is expected to possess a good thermal stability in addition to ensured accuracy and extreme sensitivity

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Summary

Introduction

Recent research in DNA biosensor fabrication has gained remarkable progress by offering reliable approaches to quantify DNA and detect mutations [1,2,3]. Coating the electrode with conductive polymer, such as polyaniline or PEDOT, has become a prospective strategy to construct a biosensor for DNA quantification because the conductive film creates a unique surface morphology to immobilize the DNA probe and has excellent electrochemical aspects for detection [9,10]. Were target DNA hybridization and detection with differential pulse voltammetry (DPV) were similar to our similar to report our previous report [17]. Of target DNA, (6) daunorubicin hydrochloride (DNM) intercalation and electrochemical detection. Hybridization of target DNA, (6) daunorubicin hydrochloride (DNM) intercalation and (B) Proposed scheme for PCR product detection: (1) PCR reaction, (2) heating and cooling procedure electrochemical detection. Working electrode in response to the concentration of target DNA

Chemicals
Pt Electrode Modification
Designs of Probe and Target DNA
DNA Biosensor Fabrication and Performance
Results and Discussion
DNA Biosensor Measurement
Conclusions
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