Electrochemical Detection of DNA in Agriculture Using Lamp-Based Amplification

• Published in: Meeting abstracts (Electrochemical Society) Vol. MA2024-02; no. 67; p. 4768
• Main Authors: Kupresanin, Ana Miodrag, Pavlovic, Zoran Miroslav, Sasic Zoric, Ljiljana, Pavlovic, Marija, Djisalov, Mila, Knezic, Teodora, Janjusevic, Ljiljana, Kanas, Nikola, Peric, Milinko, Gadjanski, Ivana
• Format: Journal Article
• Language: English
• Published: The Electrochemical Society, Inc 22.11.2024
• DOI: 10.1149/MA2024-02674768mtgabs
• ISSN: 2151-2043

The variety and complexity of genetically modified organisms (GMOs) are increasing, which makes their detection harder than ever before. This impacts the traceability, safety, and monitoring of food. Current research indicates that GM crop technology may pose risks to human health and the environment. The aim of this study is the development of a LAMP-based GMO electrochemical detection device. LAMP amplification of P-35S, P-FMV, and T-NOS regions (three elements that are most often present in genetically modified plants as part of the transgenic construct) is done using gBlocks, synthetic dsDNA fragments that have an identical DNA sequence to that of the targeted part of a transgenic construct. Using single-stranded DNA probes containing methylene blue as a redox probe, the electrochemical DNA sensor detects LAMP products that are complementary to the target DNA. The probes were attached to the gold electrodes functionalized by gold nanoparticles, and 2D nanomaterials (MoS 2 , Mxenes, and reduced graphene oxide). The electrochemical methods, such as alternating current voltammetry, square wave voltammetry, and differential pulse voltammetry were employed in testing the electrochemical detection response of the DNA probes immobilized on 2D nanomaterials. The research has shown that the obtained DNA detection signal depends significantly on the methods (and setup parameters) used in electrochemical detection, and the nanomaterials used on electrodes. The electrochemical detection resulted in a signal decrease consequential to the conformational changes of the DNA probes upon attachment of the target DNA. The methylene blue molecules attached at the top of the DNA probes get higher resistance in the electron transfer to the electrode materials, which makes the target DNA detection quantitatively and qualitatively visible. These results are promising for the development of a device that will enable a specific, sensitive, fast, cost-effective, and precise in-field detection system for the routine detection of GMOs. **Acknowledgments: *This project is supported by The Science Fund of the Republic of Serbia DEVELOPMENT Program - Green program of cooperation between science and industry - LABOUR project (grant agreement No. 6710), and The Ministry of Science, Technological Development and Innovation of the Republic of Serbia (451-03-66/2024-03/200358). *This work is supported through the ANTARES project that has received funding from the European Union's Horizon 2020 research and innovation program under grant agreement SGA-CSA. No. 739570 under FPA No. 664387. https://doi.org/10.3030/739570 Figure 1