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PLoS Negl Trop Dis . Development and evaluation of a rapid and simple diagnostic assay for COVID-19 based on loop-mediated isothermal amplification

tetano

Editor, Senior Moderator
PLoS Negl Trop Dis


. 2020 Nov 4;14(11):e0008855.
doi: 10.1371/journal.pntd.0008855. Online ahead of print.
Development and evaluation of a rapid and simple diagnostic assay for COVID-19 based on loop-mediated isothermal amplification


Rokusuke Yoshikawa[SUP] 1 2 [/SUP], Haruka Abe[SUP] 2 [/SUP], Yui Igasaki[SUP] 2 [/SUP], Saeki Negishi[SUP] 3 [/SUP], Hiroaki Goto[SUP] 3 [/SUP], Jiro Yasuda[SUP] 1 2 4 [/SUP]



Affiliations

Abstract

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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