Robène I. auteur aut IRD Jouen E. auteur aut IRD Maillot-Lebon V. auteur aut IRD Fenelon B. T. auteur aut IRD Hascoat J. auteur aut IRD Pecrix Y. auteur aut IRD Richard Damien auteur aut IRD Jaffar-Bandjee M. C. auteur aut IRD Mavingui P. auteur aut IRD Chiroleu F. auteur aut IRD Becker N. auteur aut IRD Poubeau P. auteur aut IRD Ramiandrisoa M. auteur aut IRD Sin M. auteur aut IRD Costet L. auteur aut IRD Laurent A. auteur aut IRD Laurent P. auteur aut IRD Chabirand A. auteur aut IRD Moreau A. auteur aut IRD Reynaud B. auteur aut IRD Jeuffrault E. auteur aut IRD Cetre-Sossah C. auteur aut IRD text journalArticle eng text/pdf reformatted digital access 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. specialized SARS-CoV-2 triplex-RT-LAMP detection point-of-care testing 020 052 10 1 bpaf010 [16 p.] 2025 https://www.documentation.ird.fr/hor/fdi:010092872 10.1093/biomethods/bpaf010 [F B010092872] https://www.documentation.ird.fr/hor/fdi:010092872 https://horizon.documentation.ird.fr/exl-doc/pleins_textes/2025-04/010092872.pdf IRD - Base Horizon / Pleins textes 2025-04-02 2025-11-04 fdi:010092872 fre