Duplex fluorescence reverse transcription loop-mediated isothermal amplification on a partitioned nanofluidic chip

Over the past decades, the frequency of viral outbreaks has increased substantially worldwide, driven in part by global travel and resulting in millions of deaths each year. This trend underscores the urgent need for rapid, simple, and accessible diagnostic tools for infectious disease detection. Here, we present a partitioned nanofluidic chip for detecting purified, in vitro-transcribed RNA targets, delivering results within 55 minutes at a cost of less than $0.50 per chip. The nanofluidic chip employs reverse transcription loop-mediated isothermal amplification (RT-LAMP) for sensitive and specific target amplification. Reaction reagents are compartmentalized into numerous nanofluidic reservoirs, enabling quantitative detection while minimizing contamination risks. Using a single chip, we successfully detect both SARS-CoV-2 and Influenza H3 RNA with a concentration of 727.2 aM (∼437 copies/µL), demonstrating the partitioned nanofluidic chip’s potential as a rapid, low-cost, and scalable diagnostic platform for timely outbreak control.