[HTML][HTML] Development and evaluation of a rapid and simple diagnostic assay for COVID-19 based on loop-mediated isothermal amplification

R Yoshikawa, H Abe, Y Igasaki, S Negishi… - PLoS neglected …, 2020 - journals.plos.org
R Yoshikawa, H Abe, Y Igasaki, S Negishi, H Goto, J Yasuda
PLoS neglected tropical diseases, 2020journals.plos.org
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 …
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 103 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.
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