期刊
NATURE BIOMEDICAL ENGINEERING
卷 6, 期 8, 页码 944-+出版社
NATURE PORTFOLIO
DOI: 10.1038/s41551-022-00917-y
关键词
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资金
- Shurl & Kay Curci Foundation
- National Institutes of Health (NIH)
- DARPA [N66001-20-2-4033]
- NIH [R01GM131073, DP5OD021369, R01GM127463]
- NSF Graduate Research Fellowship
- Emergent Ventures
This article reports a COVID-19 testing method implemented in a microfluidic device, which can rapidly detect SARS-CoV-2 RNA in saliva samples using isothermal RNA amplification and Cas13-mediated cleavage of a quenched fluorophore. The method has high sensitivity and specificity, and the testing can be completed within 60 minutes.
A COVID-19 test implemented in an automated microfluidic device and leveraging isothermal RNA amplification followed by T7 transcription and Cas13-mediated cleavage of a quenched fluorophore rapidly detects SARS-CoV-2 RNA in saliva samples. Rapid nucleic acid testing is central to infectious disease surveillance. Here, we report an assay for rapid COVID-19 testing and its implementation in a prototype microfluidic device. The assay, which we named DISCoVER (for diagnostics with coronavirus enzymatic reporting), involves extraction-free sample lysis via shelf-stable and low-cost reagents, multiplexed isothermal RNA amplification followed by T7 transcription, and Cas13-mediated cleavage of a quenched fluorophore. The device consists of a single-use gravity-driven microfluidic cartridge inserted into a compact instrument for automated running of the assay and readout of fluorescence within 60 min. DISCoVER can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in saliva with a sensitivity of 40 copies mu l(-1), and was 94% sensitive and 100% specific when validated (against quantitative PCR) using total RNA extracted from 63 nasal-swab samples (33 SARS-CoV-2-positive, with cycle-threshold values of 13-35). The device correctly identified all tested clinical saliva samples (10 SARS-CoV-2-positive out of 13, with cycle-threshold values of 23-31). Rapid point-of-care nucleic acid testing may broaden the use of molecular diagnostics.
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