Integrating LAMP-coupled Modification SPCE with SiNWs/PtNPs in an Electrochemical DNA Biosensor for Real-time Monitoring of Porcine DNA Amplification

This study presents a novel loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of porcine DNA, addressing critical needs in food safety and compliance with halal and kosher dietary laws. The key innovation lies in the design of highly specific primers targeting Sus scrofa mitochondrial DNA, ensuring accurate identification with minimal cross-reactivity. The LAMP technique was selected for its rapid amplification, isothermal operation, and simplified equipment requirements, making it suitable for on-site applications and reducing overall costs. Optimization of reaction conditions enabled reliable DNA amplification at 63°C within 60 minutes. The progress of the amplification was monitored in real time by measuring turbidity, while successful DNA synthesis was further confirmed by gel electrophoresis. For sensitive detection and quantification, an electrochemical DNA biosensor was integrated into the workflow. This biosensor utilizes a silicon nanowire–platinum nanoparticle–modified screen-printed carbon electrode and ferrocenylnaphthalene diimide as a dsDNA intercalator, enabling precise, real-time monitoring of the LAMP reaction. The optimized assay achieved a detection limit of 175.2 ng/μL, demonstrating high sensitivity for low-level porcine DNA detection. Specificity was validated across various meat sources and processed foods, with no false positives observed. The biosensor-based method also successfully detected porcine DNA in mixed meat samples, highlighting its strong potential for practical use in food authenticity testing and regulatory compliance.