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Development of an electrochemical DNA biosensor to detect a foodborne pathogen


Nordin, Noordiana and Yusof, Nor Azah and Radu, Son and Hushiarian, Roozbeh (2018) Development of an electrochemical DNA biosensor to detect a foodborne pathogen. Journal of Visualized Experiments (136). pp. 1-17. ISSN 1940-087X


Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that contributes to a large proportion of public health problems globally, significantly affecting the rate of human mortality and morbidity. Conventional methods for the detection of V. parahaemolyticus such as culture-based methods, immunological assays, and molecular-based methods require complicated sample handling and are time-consuming, tedious, and costly. Recently, biosensors have proven to be a promising and comprehensive detection method with the advantages of fast detection, cost-effectiveness, and practicality. This research focuses on developing a rapid method of detecting V. parahaemolyticus with high selectivity and sensitivity using the principles of DNA hybridization. In the work, characterization of synthesized polylactic acid-stabilized gold nanoparticles (PLA-AuNPs) was achieved using X-ray Diffraction (XRD), Ultraviolet-visible Spectroscopy (UV-Vis), Transmission Electron Microscopy (TEM), Field-emission Scanning Electron Microscopy (FESEM), and Cyclic Voltammetry (CV). We also carried out further testing of stability, sensitivity, and reproducibility of the PLA-AuNPs. We found that the PLA-AuNPs formed a sound structure of stabilized nanoparticles in aqueous solution. We also observed that the sensitivity improved as a result of the smaller charge transfer resistance (Rct) value and an increase of active surface area (0.41 cm2). The development of our DNA biosensor was based on modification of a screen-printed carbon electrode (SPCE) with PLA-AuNPs and using methylene blue (MB) as the redox indicator. We assessed the immobilization and hybridization events by differential pulse voltammetry (DPV). We found that complementary, non-complementary, and mismatched oligonucleotides were specifically distinguished by the fabricated biosensor. It also showed reliably sensitive detection in cross-reactivity studies against various food-borne pathogens and in the identification of V. parahaemolyticus in fresh cockles.

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Additional Metadata

Item Type: Article
Divisions: Institute of Advanced Technology
Institute of Tropical Agriculture and Food Security
DOI Number: https://doi.org/10.3791/56585
Publisher: Journal of Visualized Experiments
Keywords: Vibrio parahaemolyticus; Electrochemical DNA biosensor; Foodborne pathogen
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 26 Apr 2020 04:29
Last Modified: 26 Apr 2020 04:29
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.3791/56585
URI: http://psasir.upm.edu.my/id/eprint/72251
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