Abstract
Biosensors are transforming point-of-care diagnostics by simplifying the detection process and enabling rapid, accurate testing. This study introduces a novel, reusable biosensor designed for direct viral RNA detection from unfiltered saliva, targeting SARS-CoV-2. Unlike conventional methods requiring filtration, our biosensor leverages a unique electrode design that prevents interference from saliva debris, allowing precise measurements. The biosensor is based on electrochemical principles, employing oligonucleotide probes immobilized on a hydrophobic-coated electrode, which prevents air bubbles and salt crystal formation. During validation, the biosensor demonstrated a sensitivity and specificity of 100%, accurately identifying SARS-CoV-2 in saliva samples without false positives or negatives. Cross-validation with RT-qPCR, the gold standard for COVID-19 diagnostics, confirmed the reliability of our device. The biosensor's performance was tested on 60 participants, yielding 12 true positive results and 48 true negatives, aligning perfectly with RT-qPCR outcomes. This reusable, easy-to-use biosensor offers significant potential for point-of-care applications in various healthcare settings, providing a fast, efficient, and cost-effective method for detecting viral infections such as COVID-19. Its robust design, minimal sample preparation requirements, and multiple-use capability mark a significant advancement in biosensing technology.