Viral Infection-Inspired Autonomous Detection of Fusion-Competent Viruses for Screening and Environmental Surveillance

Abstract

The persistent burden of respiratory viruses requires rapid, simple, and robust screening and environmental surveillance technologies that enable widespread and frequent testing. Importantly, these technologies should be based on infectivity-relevant signals, as RNA detection alone has limited correlation with transmission risk. Here, we present a membrane fusion-mediated platform that autonomously detects viruses by recapitulating the native viral entry mechanism. Fusogenic vesicles selectively fuse with fusion-competent viral particles, triggering encapsulated CRISPR-Cas13a components to generate fluorescent signals upon recognition of the released viral RNA. Through an autonomous workflow and accelerated signal generation within a confined vesicle, our platform achieves one-step detection of viruses within 2 min. The assay robustly detects three major respiratory viruses, with analytical sensitivities down to 5 TCID50/mL for RSV and 50 TCID50/mL for SARS-CoV-2 and IAV. Clinical validation with 100 nasopharyngeal samples achieved 91.7% sensitivity. Remarkably, the sprayable format enables large-area surveillance of surface contamination—like luminol revealing hidden bloodstains, it makes invisible viral threats visible. This approach establishes an intuitive real-time detection platform, extending beyond clinical specimens to encompass environmental threats.