Figure 1. Design Process diagram for different cyclical stages of our product’s development.  

Figure 2.  This figure above demonstrates the higher level scope of our biosensor assay. 

Figure 3. This graphic shows the three main steps of our electrochemical protocol: 1) The functionalization of electrodes surface. 2) The blocking step via elimination of non-specific DNA binding.  3) Target DNA hybridization on electrode surface. 

 Figure 5. The circuit used for the 3-electrode system indicating the counter electrode (Green) and Reference Electrode (Blue) with a potentiostat (PalmSens). 

Testing 

Accuracy and Verification

The development and optimization of our electrochemical biosensor have underscored its accuracy in detecting specific DNA sequences, including the D614G mutation. This precision is crucial for the effective monitoring and management of infectious diseases. To further enhance our confidence in the biosensor's specificity and sensitivity, an off-chip verification method using SYBR Green is currently being pursued. This approach involves fluorescent dye binding to double-stranded DNA, illuminating the successful hybridization events and thus serving as an independent validation of the on-chip results. This additional verification step is designed to eliminate any ambiguity regarding hybridization efficiency and specificity, thereby reinforcing the biosensor's accuracy. The incorporation of SYBR Green, known for its high affinity to double-stranded DNA and its capability to provide rapid and reliable confirmation of molecular interactions, represents a significant stride towards establishing our biosensor as a robust tool for molecular diagnostics.

Relevance and Implementation Potential

Our electrochemical biosensor demonstrates a significant potential for practical implementation, offering a timely and relevant solution to the current needs in infectious disease diagnostics, particularly for COVID-19 and its mutations. The biosensor's design allows for the straightforward incorporation of multiple probes, enabling the detection of various COVID-19 mutations within a single assay. This multiplexing capability is not only efficient but also cost-effective, making it an invaluable tool for large-scale screening and monitoring efforts. The testing results have validated the biosensor's functionality and reliability, indicating that its implementation in clinical and public health settings is not only viable but also highly beneficial. Its ability to provide rapid, accurate, and specific detection of viral RNA sequences positions it as a critical asset in the ongoing fight against the pandemic, offering a scalable solution that can adapt to the evolving landscape of COVID-19 variants.

Comparison to Alternatives

When compared to alternative diagnostic approaches such as health patches or sweat-based airborne disease detection biosensors, our electrochemical biosensor offers distinct advantages. While health patches and sweat-based sensors provide non-invasive monitoring capabilities, they often lack the specificity and sensitivity required for accurate viral detection, particularly in the early stages of infection. Our electrochemical biosensor, on the other hand, directly targets the genetic markers of the virus, providing a level of precision that is critical for early diagnosis and effective disease management. Furthermore, the electrochemical biosensor's rapid response time and the potential for real-time analysis surpass the slower processing times associated with alternative methods. This immediacy is crucial for timely decision-making in clinical settings, where rapid turnaround times can significantly impact patient outcomes.

Leaders: Lidia & Riam