Performing point-of-care molecular testing for SARS-CoV-2 with RNA extraction and isothermal amplification
In: ISSN: 1932-6203, 2021
Online
academicJournal
Zugriff:
International audience ; To respond to the urgent need for COVID-19 testing, countries perform nucleic acid amplification tests (NAAT) for the detection of SARS-CoV-2 in centralized laboratories. Real-time RT—PCR (Reverse transcription—Polymerase Chain Reaction), used to amplify and detect the viral RNA., is considered, as the current gold standard for diagnostics. It is an efficient process, but the complex engineering required for automated RNA extraction and temperature cycling makes it incompatible for use in point of care settings [1]. In the present work, by harnessing progress made in the past two decades in isothermal amplification and paper microfluidics, we created a portable test, in which SARS-CoV-2 RNA is extracted, amplified isothermally by RT—LAMP (Loop-mediated Isothermal Amplification), and detected using intercalating dyes or fluorescent probes. Depending on the viral load in the tested samples, the detection takes between twenty minutes and one hour. Using a set of 16 pools of naso-pharyngal swab eluates, we estimated a limit of detection comparable to real-time RT-PCR (i.e. 1 genome copies per microliter of clinical sample) and no cross‐reaction with eight major respiratory viruses currently circulating in Europe. We designed and fabricated an easy-to-use portable device called “COVIDISC” to carry out the test at the point of care. The low cost of the materials along with the absence of complex equipment will expedite the widespread dissemination of this device. What is proposed here is a new efficient tool to help managing the pandemics.
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Performing point-of-care molecular testing for SARS-CoV-2 with RNA extraction and isothermal amplification
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Autor/in / Beteiligte Person: | Garneret, Pierre ; Coz, Etienne ; Martin, Elian ; Manuguerra, Jean-Claude ; Brient-Litzler, Elodie ; Enouf, Vincent ; González Obando, Daniel Felipe ; Olivo-Marin, Jean-Christophe ; Monti, Fabrice ; van der Werf, Sylvie ; Vanhomwegen, Jessica ; Tabeling, Patrick ; Chimie-Biologie-Innovation (UMR 8231) (CBI) ; Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris) ; Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS) ; Institut Pierre-Gilles de Gennes pour la Microfluidique (IPGG) ; École normale supérieure - Paris (ENS-PSL) ; Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP) ; Université Paris Sciences et Lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris) ; Université Paris Sciences et Lettres (PSL)-Institut Curie Paris -Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS) ; Cellule d'Intervention Biologique d'Urgence (Centre National de Référence) - Laboratory for Urgent Response to Biological Threats (National Reference Center) (CIBU) ; Environnement et Risques infectieux - Environment and Infectious Risks (ERI) ; Institut Pasteur Paris (IP)-Institut Pasteur Paris (IP) ; Institut Pasteur Paris (IP) ; Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)) ; Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité) ; Centre National de Référence des virus des infections respiratoires (dont la grippe) - National Reference Center Virus Influenzae Paris (CNR - laboratoire coordonnateur) ; Analyse d'images biologiques - Biological Image Analysis (BIA) ; Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS) ; Laboratoire Microfluidique, MEMS, Nanostructures (MMN) ; Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Centre National de la Recherche Scientifique (CNRS) ; IPGG platform (ANR-10-EQPX-34) (PT,PG,EC,EM), Carnot Pasteur Maladies Infectieuses program (ANR 11-CARN 017-01) (JV,JCM), ANR-20-COVI-000 (EB,JV,JCM,PT,EM,C), FONDS ESPCI (EC) ; ANR-10-EQPX-0034,IPGG,Institut Pierre Gilles de Gennes pour la microfluidique(2010) |
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Zeitschrift: | ISSN: 1932-6203, 2021 |
Veröffentlichung: | HAL CCSD ; Public Library of Science, 2021 |
Medientyp: | academicJournal |
DOI: | 10.1371/journal.pone.0243712 |
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