Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly pathogenic novel coronavirus that has caused a worldwide outbreak. Here we describe a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay that uses a portable device for efficient detection of SARS-CoV-2. This RT-LAMP assay specifically detected SARS-CoV-2 without cross-reacting with the most closely related human coronavirus, SARS-CoV. Clinical evaluation of nasal swab samples from suspected SARS-CoV-2 pneumonia (COVID-19) patients showed that the assay could detect over 23.7 copies within 15 min with a 100% probability. Since the RT-LAMP assay can be performed with a portable battery-supported device, it is a rapid, simple, and sensitive diagnostic assay for COVID-19 that can be available at point-of-care. We also developed the RT-LAMP assay without the RNA extraction step–Direct RT-LAMP, which could detect more than 1.43 x 10 3 copies within 15 min with a 100% probability in clinical evaluation test. Although the Direct RT-LAMP assay was less sensitive than the standard RT-LAMP, the Direct RT-LAMP assay can be available as the rapid first screening of COVID-19 in poorly equipped areas, such as rural areas in developing countries. Author summary Rapid, accurate and simple diagnostic assay is required to provide an appropriate treatment and establish a surveillance system to prevent the spread of COVID-19. In this study, we developed the RT-LAMP assay using a portable platform equipped with a battery for detection of SARS-CoV-2 in clinical specimens. It could detect over 23.7 copies within 15 min. Since RNA extraction step is one of the main obstacles for rapid diagnosis, we also developed the RT-LAMP assay without this step, Direct RT-LAMP. This assay could detect over 203 copies of viral RNA in virus-spiked swab solution within 10.5 min. Since our RT-LAMP assay is a rapid, simple and highly sensitive diagnostic system for identifying COVID-19 case, it would be useful for the detection of SARS-CoV-2, especially in the field or the settings with insufficient infrastructure.
【초록키워드】 COVID-19, Treatment, severe acute respiratory syndrome coronavirus 2, SARS-CoV-2, coronavirus, Pneumonia, SARS-CoV, diagnostic, Diagnosis, severe acute respiratory syndrome Coronavirus, Novel coronavirus, RNA extraction, Probability, point-of-care, RT-LAMP, Viral, Surveillance, Loop-mediated isothermal amplification, clinical evaluation, outbreak, clinical, Patient, Rapid, reverse transcription, isothermal amplification, Viral RNA, Swab, nasal swab, SARS-CoV-2 pneumonia, respiratory, specimens, platform, rural areas, acute respiratory syndrome, acute respiratory syndrome coronavirus, battery, COVID-19 case, highly pathogenic, Direct, Prevent, spread of COVID-19, human coronavirus, performed, detect, caused, required, less, 【제목키워드】 COVID-19, diagnostic, Loop-mediated isothermal amplification, isothermal amplification, development,