Washing-free Electrochemical Strategy to Detect Target DNA Utilizing Peroxidase Mimicking DNAzyme
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pISSN 1226-8372 eISSN 1976-3816
RESEARCH PAPER
Washing-free Electrochemical Strategy to Detect Target DNA Utilizing Peroxidase Mimicking DNAzyme Sang Mo Lee, Sujeong Shin, Hyo Yong Kim, Byoung Yeon Won, Jun Ki Ahn, Ki Soo Park, and Hyun Gyu Park
Received: 24 August 2020 / Revised: 14 September 2020 / Accepted: 15 September 2020 © The Korean Society for Biotechnology and Bioengineering and Springer 2020
Abstract We herein describe a novel washing-free electrochemical strategy for target DNA detection by utilizing the peroxidase mimicking DNAzyme. The DNAzymeincorporated DNA probes, the key components of this strategy, are designed to be not able to form G-quadruplex structure in the initial state such that the peroxidase mimicking activity of DNAzyme is kept suppressed. The DNAzyme sequences, however, would be converted to the catalytically active G-quadruplex structure by the presence of target DNA, whose peroxidase mimicking activity then promote the precipitation reaction of 4-chloronaphthol (4CN) on the electrode. Due to the precipitates produced on the electrode surface, the electrochemical reaction between the redox materials and electrode surface is inhibited, consequently leading to a significant increase of the impedance signal. Based on this novel electrochemical design principle, we successfully detected the target DNA from sexually transmitted disease (STD) pathogens such as Sang Mo Lee†, Sujeong Shin†, Hyo Yong Kim, Byoung Yeon Won, Jun Ki Ahn, Hyun Gyu Park* Department of Chemical and Biomolecular Engineering (BK 21+ program), KAIST, Daejeon 34141, Korea Tel: +82-42-350-3932; Fax: +82-42-350-3910 E-mail: [email protected] Sujeong Shin Division of Safety Analysis, National Agricultural Products Quality Management Service, Ministry of Agriculture, Food and Rural Affairs, Gimcheon 39660, Korea Byoung Yeon Won Boditech Med Inc., Chuncheon 24398, Korea Jun Ki Ahn Human Convergence Technology Group, Korea Institute of Industrial Technology (KITECH), Ansan 15588, Korea Ki Soo Park Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Korea †
These authors equally contributed to this work.
Chlamydia trachomatis, Trichomonas vaginalis, and Herpes simplex virus type 2 in real patient samples, verifying its practical diagnostic capability in the clinical applications. Keywords: peroxidase mimicking DNAzyme, target DNA detection, electrochemical sensor, 4-chloronaphthol, sexually transmitted disease
1. Introduction Target DNA detection has been of great significance due to its wide utility in the area of human genetics, forensic science, and molecular diagnostics [1-3]. The quantitative PCR (qPCR) is currently used as a gold standard method for the identification of target DNA, which follows the kinetics of the PCR and detects PCR products during the process of amplification. With qPCR, accurate and reproducible quantification of pathogen concentration could be incorporated into the amplification process and qPCR is now widely employed to identify the target DNA from
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