The COVID-19 pandemic caused by the SARS-CoV-2 virus, which first emerged in December 2019, represents an ongoing global public health emergency. Here, we developed an improved and highly sensitive approach to SARS-CoV-2 detection via coupling bioluminescence in real-time (BART) and reverse-transcriptase loop-mediated amplification (RT-LAMP) protocols (RT-LAMP-BART) and was also compatible with a digital LAMP system (Rainsuit), which did not allow for real-time quantification but did, nonetheless, facilitate absolute quantification with a comparable detection limit of 10 4 copies/mL. Through improving RNA availability in samples to ensure the target RNA present in reaction, we additionally developed a simulated digital RT-LAMP approach using this same principle to enlarge the overall reaction volume and to achieve real-time detection with a limit of detection of 10 copies/mL, and with further improvements in the overall dynamic range of this assay system being achieved through additional optimization.
【저자키워드】 SARS-CoV-2, accurate and rapid identification, RT-LAMP-BART, digital PCR system, digital RT-LAMP approach, 【초록키워드】 protocol, COVID-19 pandemic, SARS-CoV-2 virus, RNA, improvement, amplification, RT-LAMP, SARS-CoV-2 detection, LAMP, limit of detection, bioluminescence, quantification, Detection limit, dynamic range, Volume, reaction, global public health, approach, caused, facilitate, comparable, the SARS-CoV-2 virus, 【제목키워드】 detection, RNA, PCR, Digital, novel, ORF1ab, coupling, the SARS-CoV-2,