RUNCOV: a one-pot triplex real-time RT-LAMP as a point-of-care diagnostic tool for detecting SARS-CoV-2

被引：1

作者：

Robene, Isabelle ^{[1
]}

Jouen, Emmanuel ^{[1
]}

Maillot-Lebon, Veronique ^{[1
]}

Fenelon, Babbitha T. ^{[1
]}

Hascoat, Jeremy ^{[1
]}

Pecrix, Yann ^{[1
]}

Richard, Damien ^{[1
]}

Jaffar-Bandjee, Marie-Christine ^{[2
]}

Mavingui, Patrick ^{[3
,4
]}

Chiroleu, Frederic ^{[1
]}

Becker, Nathalie ^{[5
]}

Poubeau, Patrice ^{[6
]}

Ramiandrisoa, Mahery ^{[7
]}

Sin, Michel ^{[7
]}

Costet, Laurent ^{[1
]}

Laurent, Annie ^{[8
]}

Laurent, Philippe ^{[9
]}

Chabirand, Aude ^{[10
]}

Moreau, Aurelie ^{[10
]}

Reynaud, Bernard ^{[1
]}

Jeuffrault, Eric ^{[1
]}

Cetre-Sossah, Catherine ^{[11
]}

机构：

[1] CIRAD, UMR PVBMT, 7 chemin lIRAT,Ligne Paradis, F-97410 St Pierre, La Reunion, France

[2] Ctr Hosp Univ CHU Reun Felix Guyon, Serv Microbiol, Sect Virol, F-97400 St Denis, La Reunion, France

[3] Univ Reunion, UMR Proc Infect Milieu Insulaire Trop PIMIT, CNRS 9192, INSERM 1187,IRD 249, F-97490 St Clotilde, La Reunion, France

[4] Plateforme Technol CYROI, F-97490 Sainte Clotilde, La Reunion, France

[5] Sorbonne Univ, Univ Antilles, Inst Systemat Evolut Biodivers ISYEB, Museum Natl dHist Naturelle,CNRS,EPHE, F-75005 Paris, France

[6] Ctr Hosp Univ CHU La Reunion, Serv Malad Infectieuses, South Site, F-97410 St Pierre, La Reunion, France

[7] Labs Anal Med Cerballiance Reunion, F-97420 St Pierre, La Reunion, France

[8] Ctr Hosp Univ CHU Reunion, ART Ctr Med Assistance Reprod, South site, F-97410 St pierre, La Reunion, France

[9] Univ Reunion, UMR PVBMT, F-97410 St Pierre, La Reunion, France

[10] Trop Pests & Pathogens RAPT unit, Plant Hlth Lab, French Agcy Food Environm & Occupat Hlth & Safety, F-97410 St Pierre, La Reunion, France

[11] CIRAD, UMR ASTRE, F-97490 St Clotilde, La Reunion, France

来源：

BIOLOGY METHODS & PROTOCOLS | 2025年 / 10卷 / 01期

关键词：

SARS-CoV-2; triplex-RT-LAMP; detection; point-of-care testing; MEDIATED ISOTHERMAL AMPLIFICATION; SINGLE; PCR;

D O I：

10.1093/biomethods/bpaf010

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 R & eacute;union in different contexts outside the laboratory.

引用

页数：16

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