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The current gold standard for COVID-19 confirmation is RT-PCR, which possesses high sensitivity and specificity, but a long turnaround time has been a drawback. Here, we introduce a nanoPCR platform that can yield breakthroughs in RT-PCR speed and decentralize the diagnostic testing. A key innovation is to exploit magneto-plasmonic nanoparticles for dual purposes: fast thermocycling through plasmonic heating and enhanced signal detection via magnetic fluorescence switch. We further integrated these functionalities into a portable system to streamline point-of-care (POC) COVID-19 diagnostics with a “one-step-finish” approach from sampling to result. When applied with clinical COVID-19 samples, nanoPCR achieved an analytical performance of sensitivity and specificity comparable to that of a benchtop instrument but completed the entire detection within 18 minutes (vs. 2.5 hours for regular RT-PCR), which makes it ideal for rapid POC diagnostics. This platform technology is readily scalable for higher throughput and expandable to other disease targets, promising broader impacts in on-site molecular testing.

Tetrahydrocannabinol (THC), the primary psychoactive ingredient of cannabis, impairs cognitive and motor function in a concentration-dependent fashion. Drug testing is commonly performed for employment and law enforcement purposes; however, available tests produce low-sensitive binary results (lateral flow assays) or have long turnaround (gas chromatography-mass spectrometry). To enable on-site THC quantification in minutes, we developed a rapid assay for oral THC analysis called EPOCH, express probe for on-site cannabis inhalation. EPOCH features distinctive sensor design such as a radial membrane and transmission optics, all contained in a compact cartridge. This integrated approach permitted assay completion within 5 minutes with a detection limit of 0.17 ng/mL THC, which is below the regulatory guideline (1 ng/mL). As a proof of concept for field testing, we applied EPOCH to assess oral fluid samples from cannabis users (n = 43) and controls (n = 43). EPOCH detected oral THC in all specimens from cannabis smokers (median concentration, 478 ng/mL) and THC-infused food consumers. Longitudinal monitoring showed a fast drop in THC concentrations within the first 6 hours of cannabis smoking (half-life, 1.4 hours).

Prompt diagnosis, patient isolation, and contact tracing are key measures to contain the coronavirus disease 2019 (COVID-19). Molecular tests are the current gold standard for COVID-19 detection, but are carried out at central laboratories, delaying treatment and control decisions. Here we describe a portable assay system for rapid, onsite COVID-19 diagnosis. Termed CODA (CRISPR Optical Detection of Anisotropy), the method combined isothermal nucleic acid amplification, activation of CRISPR/Cas12a, and signal generation in a single assay, eliminating extra manual steps. Importantly, signal detection was based on the ratiometric measurement of fluorescent anisotropy, which allowed CODA to achieve a high signal-to-noise ratio. For point-of-care operation, we built a compact, standalone CODA device integrating optoelectronics, an embedded heater, and a microcontroller for data processing. The developed system completed SARS-CoV-2 RNA detection within 20 min of sample loading; the limit of detection reached 3 copy/μL. When applied to clinical samples (10 confirmed COVID-19 patients; 10 controls), the rapid CODA test accurately classified COVID-19 status, in concordance with gold-standard clinical diagnostics., Highlights • We developed a CODA (CRISPR Optical Detection of Anisotropy) system for point-of-care COVID-19 diagnostics. • CODA detected SARS-CoV-2 RNA down to 3 copy/μL within 20 min in a one-pot assay. • CODA assays accurately identified COVID-19 when applied to patient samples (n = 20).