Fusogenic-Reactor-Based Lateral Flow Assay for Extraction-free, Multiplexed Viral RNA Detection

Descrizione del progetto

Test multiplex domestico per il rilevamento dei virus

I test multiplex in grado di rilevare simultaneamente un certo numero di virus, come l’influenza A e il SARS-CoV-2, sono essenziali per un’efficace sorveglianza della malattia. Gli attuali diagnostici virali per uso domestico mancano di accuratezza, mentre i test basati sulla PCR sono laboriosi e richiedono risorse specializzate. Con il sostegno del programma di azioni Marie Skłodowska-Curie, il progetto FusoLipo-LFA svilupperà un test innovativo basato su vescicole lipidiche che imitano le cellule e possono fondersi con i virus. Queste vescicole contengono speciali strumenti molecolari in grado di rilevare l’RNA virale e di amplificare il segnale, facilitando la rilevazione anche di poche particelle virali. Questo approccio supera le complesse fasi di estrazione e offre un test domestico rapido, accurato e facile da usare per diversi virus.

Obiettivo

In this fellowship, I will leverage recent advances in membrane-fusion-based biosensor technologies to develop an extraction-free, multiplexed lateral flow assay (LFA) for rapid, home-based viral RNA detection. The COVID-19 pandemic exposed critical limitations in virus diagnostics, with antigen-based kits lacking sensitivity and specificity, while RT-qPCR tests, though accurate, are slow, resource-intensive, and impractical for widespread deployment—highlighting the urgent need for field-deployable molecular assays that deliver reliable results. Given the coexistence of multiple respiratory viruses, multiplexed detection is also crucial. This project aims to combine my expertise in membrane-fusion-based biosensor design with Professor Dame Molly Stevens' (University of Oxford) expertise in LFA-based biosensors to create a multiplex LFA system for viral RNA detection using membrane fusion approach. The system will first employ cell-mimetic fusogenic lipid vesicles (FusoLipo) encapsulating CRISPR toolkits, facilitating viral membrane fusion, RNA capture, and CRISPR-mediated detection. By utilizing two types of FusoLipo, the platform can simultaneously detect influenza A (H1N1) and SARS-CoV-2 without the need for complex RNA extraction, minimizing contamination risks. Once the vesicle’s cargo is applied to the LFA, catalytic nanoparticles (Nanozyme) will amplify the signal at the detection bands, increasing sensitivity. This platform ultimately provides a accurate, and user-friendly solution for home-based multiplexed viral RNA detection. Through this fellowship, I will gain access to specialized expertise and cutting-edge facilities in lipid vesicle formulation, molecular technology, LFA development, and inorganic materials synthesis in the Stevens group, alongside advanced instrumentation to characterize vesicles at the single-particle level and further study virus-FusoLipo membrane fusion, significantly advancing my capabilities in diagnostic biosensor innovation.