%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Robène, I.
%A Jouen, E.
%A Maillot-Lebon, V.
%A Fenelon, B. T.
%A Hascoat, J.
%A Pecrix, Y.
%A Richard, Damien
%A Jaffar-Bandjee, M. C.
%A Mavingui, P.
%A Chiroleu, F.
%A Becker, N.
%A Poubeau, P.
%A Ramiandrisoa, M.
%A Sin, M.
%A Costet, L.
%A Laurent, A.
%A Laurent, P.
%A Chabirand, A.
%A Moreau, A.
%A Reynaud, B.
%A Jeuffrault, E.
%A Cetre-Sossah, C.
%T RUNCOV : a one-pot triplex real-time RT-LAMP as a point-of-care diagnostic tool for detecting SARS-CoV-2
%D 2025
%L fdi:010092872
%G ENG
%J Biology Methods and Protocols
%K SARS-CoV-2 ; triplex-RT-LAMP ; detection ; point-of-care testing
%M ISI:001437414000001
%N 1
%P bpaf010 [16 ]
%R 10.1093/biomethods/bpaf010
%U https://www.documentation.ird.fr/hor/fdi:010092872
%> https://horizon.documentation.ird.fr/exl-doc/pleins_textes/2025-04/010092872.pdf
%V 10
%W Horizon (IRD)
%X Given the risk of zoonotic disease emergence, including new SARS-CoV-2 variants of COVID-19, rapid diagnostic tools are urgently needed to improve the control of the spread of infectious diseases. A one-pot triplex real-time RT-LAMP (reverse-transcription-loop-mediated isothermal amplification) assay, based on two regions of the genome SARS-CoV-2-specifically the Orf1ab and N genes-along with one internal control, the human RNase P gene, was developed. The multiplexing relies on the distinct melting peaks produced during an annealing step. This tool, named RUNCOV, was compared to the gold-standard reverse-transcription real-time quantitative PCR (RT-qPCR) assay. A simple sample preparation step was designed alongside the assay, making it ready for use on site, as a point-of-care diagnostic tool. RUNCOV is rapid (typically less than 40 minutes), highly sensitive and specific. When tested on clinical samples with known SARS-CoV-2 status, its limit of detection (LOD) ranges between 5 and 20 copies per reaction and its diagnostic sensitivity (97.44%) and specificity (100%) values are high compared to the RT-qPCR gold standard. These results were supported with an extensive in silico analysis of over 14 million genomes, demonstrating this tool was capable of detecting all known SARS-CoV-2 variants, including the most recent ones KP.3.1.1 and BA2.86.1. This molecular assay is portable, as demonstrated when it was used successfully in La Réunion in different contexts outside the laboratory.
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