The ongoing Corona virus disease (COVID-19) outbreak has become a huge global health concern. Here, we reported a novel detection platform based on the loop-mediated isothermal amplification (LAMP), termed real-time reverse transcription LAMP (rRT-LAMP) and applied it for the diagnosis of COVID-19 (COVID-19 rRT-LAMP). rRT-LAMP integrates reverse transcription, LAMP amplification, restriction endonuclease cleavage and real-time fluorescence detection into one-pot reaction, and facilitates the diagnosis of COVID-19 at 64°C for only 35 min. The ORF1ab (opening reading frame 1a/b) and NP (nucleoprotein) genes of SARS-CoV-2 were detected for diagnosing COVID-19. The limit of detection (LoD) of COVID-19 rRT-LAMP assay was 14 copies (for each marker) per vessel, and no positive results were obtained from non-SARS-CoV-2 templates. To demonstrate its feasibility, a total of 33 oropharynx swab samples collected from COVID-19 patients also were diagnosed as SARS-CoV-2 infection using COVID-19 rRT-LAMP protocol. No cross-reactivity was yielded from 41 oropharynx swab samples collected from non-COVID-19 patients. These data suggesting that the COVID-19 rRT-LAMP assay is a potential detection tool for the diagnosis of SARS-CoV-2 infection in clinical, field and disease control laboratories, and will be valuable for controlling the COVID-19 epidemic.
【저자키워드】 COVID-19, SARS-CoV-2, Loop-mediated isothermal amplification, reverse transcription, rRT-LAMP, 【초록키워드】 Corona virus, protocol, feasibility, SARS-COV-2 infection, Infection, cross-reactivity, amplification, fluorescence detection, Health, outbreak, limit of detection, cleavage, isothermal amplification, disease, platform, marker, COVID-19 epidemic, ORF1ab, COVID-19 patient, Frame, Non-COVID-19 patients, positive result, diagnosing, vessel, swab sample, collected, reported, diagnosed, applied, facilitate, diagnosis of COVID-19, diagnosis of SARS-CoV-2, 【제목키워드】 detection, novel, application,