The need for rapid, accurate, and scalable testing systems for COVID-19 diagnosis is clear and urgent. Here, we report a rapid S calable and Po rtable T esting (SPOT) system consisting of a rapid, highly sensitive, and accurate assay and a battery-powered portable device for COVID-19 diagnosis. The SPOT assay comprises a one-pot reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) followed by Pf Ago-based target sequence detection. It is capable of detecting the N gene and E gene in a multiplexed reaction with the limit of detection (LoD) of 0.44 copies/μL and 1.09 copies/μL, respectively, in SARS-CoV-2 virus-spiked saliva samples within 30 min. Moreover, the SPOT system is used to analyze 104 clinical saliva samples and identified 28/30 (93.3% sensitivity) SARS-CoV-2 positive samples (100% sensitivity if LoD is considered) and 73/74 (98.6% specificity) SARS-CoV-2 negative samples. This combination of speed, accuracy, sensitivity, and portability will enable high-volume, low-cost access to areas in need of urgent COVID-19 testing capabilities. There is a clear need for rapid, accurate and scalable Covid-19 diagnostics. Here the authors use Pf Ago to detect viral sequences amplified by RT-LAMP in a handheld battery-powered device.
【저자키워드】 Mechanical engineering, Assay systems, Biochemical assays, 【초록키워드】 SARS-CoV-2, Saliva, diagnostics, RT-LAMP, sensitivity, specificity, COVID-19 testing, Viral, Accuracy, Loop-mediated isothermal amplification, COVID-19 diagnosis, limit of detection, isothermal amplification, N gene, speed, Combination, followed by, reverse transcriptase, battery, E gene, sequence, saliva samples, viral sequences, positive sample, amplified, detect, viral sequence,