Development of an electrochemical DNA biosensor to detect a foodborne pathogen
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-bas...
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| Format: | Article |
| Language: | English |
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Journal of Visualized Experiments
2018
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| Online Access: | http://psasir.upm.edu.my/id/eprint/72251/ http://psasir.upm.edu.my/id/eprint/72251/1/Development%20of%20an%20electrochemical%20DNA%20biosensor%20to%20detect%20a%20foodborne%20pathogen.pdf |
| _version_ | 1848857073386782720 |
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| author | Nordin, Noordiana Yusof, Nor Azah Radu, Son Hushiarian, Roozbeh |
| author_facet | Nordin, Noordiana Yusof, Nor Azah Radu, Son Hushiarian, Roozbeh |
| author_sort | Nordin, Noordiana |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-15T11:51:45Z |
| format | Article |
| id | upm-72251 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T11:51:45Z |
| publishDate | 2018 |
| publisher | Journal of Visualized Experiments |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-722512020-04-26T04:29:20Z http://psasir.upm.edu.my/id/eprint/72251/ Development of an electrochemical DNA biosensor to detect a foodborne pathogen Nordin, Noordiana Yusof, Nor Azah Radu, Son Hushiarian, Roozbeh 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. Journal of Visualized Experiments 2018-06 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/72251/1/Development%20of%20an%20electrochemical%20DNA%20biosensor%20to%20detect%20a%20foodborne%20pathogen.pdf 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 https://www.jove.com/video/56585/development-an-electrochemical-dna-biosensor-to-detect-foodborne 10.3791/56585 |
| spellingShingle | Nordin, Noordiana Yusof, Nor Azah Radu, Son Hushiarian, Roozbeh Development of an electrochemical DNA biosensor to detect a foodborne pathogen |
| title | Development of an electrochemical DNA biosensor to detect a foodborne pathogen |
| title_full | Development of an electrochemical DNA biosensor to detect a foodborne pathogen |
| title_fullStr | Development of an electrochemical DNA biosensor to detect a foodborne pathogen |
| title_full_unstemmed | Development of an electrochemical DNA biosensor to detect a foodborne pathogen |
| title_short | Development of an electrochemical DNA biosensor to detect a foodborne pathogen |
| title_sort | development of an electrochemical dna biosensor to detect a foodborne pathogen |
| url | http://psasir.upm.edu.my/id/eprint/72251/ http://psasir.upm.edu.my/id/eprint/72251/ http://psasir.upm.edu.my/id/eprint/72251/ http://psasir.upm.edu.my/id/eprint/72251/1/Development%20of%20an%20electrochemical%20DNA%20biosensor%20to%20detect%20a%20foodborne%20pathogen.pdf |