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Ning, Bo; Yu, Tao; Zhang, Shengwei; Huang, Zhen; Tian, Di; Lin, Zhen; Niu, Alex; Golden, Nadia; Hensley, Krystle; Threeton, Breanna; Lyon, Christopher J; Yin, Xiao-Ming; Roy, Chad J; Saba, Nakhle S; Rappaport, Jay; Wei, Qingshan; Hu, Tony Y
Science advances, 01/2021, Volume: 7, Issue: 2Journal Article
Point-of-care COVID-19 assays that are more sensitive than the current RT-PCR (reverse transcription polymerase chain reaction) gold standard assay are needed to improve disease control efforts. We describe the development of a portable, ultrasensitive saliva-based COVID-19 assay with a 15-min sample-to-answer time that does not require RNA isolation or laboratory equipment. This assay uses CRISPR-Cas12a activity to enhance viral amplicon signal, which is stimulated by the laser diode of a smartphone-based fluorescence microscope device. This device robustly quantified viral load over a broad linear range (1 to 10 copies/μl) and exhibited a limit of detection (0.38 copies/μl) below that of the RT-PCR reference assay. CRISPR-read SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) RNA levels were similar in patient saliva and nasal swabs, and viral loads measured by RT-PCR and the smartphone-read CRISPR assay demonstrated good correlation, supporting the potential use of this portable assay for saliva-based point-of-care COVID-19 diagnosis.
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