Numerous medical conditions require extensive monitoring of diverse biomarkers, imposing a significant time and effort burden on healthcare professionals. This underscores the need for fast, reliable, and personalized point-of-care diagnostic tools. In the context of organ transplantation, the physiological status of a brain deceased organ donor requires comprehensive surveillance, accompanied by appropriate medical interventions to mitigate organ degradation. An effective strategy for monitoring this degradation involves tracking the progression and, ideally, the onset of the inflammatory response, the so-called ”cytokine storm”, by monitoring specific biomarkers: micro-RNAs. We propose two approaches based on a surface plasmon resonance imaging biosensor that combine various methodologies to amplify the hybridization signal of an oligonucleotide duplex (sandwich assay or bimaterial nanostructured biochip). In a proof-of-concept experiment involving a protein oligonucleotide complex in a sandwich assay, we successfully demonstrate signal amplification by one order of magnitude. Additionally, using the contribution of localized surface plasmons induced by nanostructuring the biochip, our simulations predict a further significant gain in amplification.
We present a Surface Plasmon Resonance Imaging (SPRI) biochip system to quantitatively detect micro-RNAs involved in the cytokine storm during an inflammatory response. The thiol composition of the self-assembled monolayer on the biochip gold surface was tuned to maximize the capture of RNAs at low concentrations. To further amplify this signal, we have developed a sandwich-like assay using oligonucleotides functionalized gold nanoparticles (AuNPs), synthesized at ambient temperature and optimized to have a high solubility in saline solutions. Sub-picomolar detection limit of those small RNAs was achieved with all these combined improvements.
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