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Development of a Fast and Affordable Diagnostic System for RNA Viruses Using Loop-Mediated Isothermal Amplification and DNA-Nanoprobe Detection
  • +5
  • Pablo Cea,
  • Sonia Arca,
  • Esperanza Gomez-Lucia,
  • Ana Domenech,
  • Mar Biarnes,
  • Angela Blanco,
  • Laura Benitez,
  • Ricardo Madrid
Pablo Cea
Complutense University of Madrid Faculty of Biological Sciences
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Sonia Arca
BioAssays, S.L.
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Esperanza Gomez-Lucia
Complutense University of Madrid Faculty of Veterinary
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Ana Domenech
Complutense University of Madrid Faculty of Veterinary
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Mar Biarnes
Centro de Sanidad Avícola de Cataluña y Aragón (CESAC)
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Angela Blanco
Centro de Sanidad Avícola de Cataluña y Aragón (CESAC)
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Laura Benitez
Complutense University of Madrid Faculty of Biological Sciences
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Ricardo Madrid
Complutense University of Madrid Faculty of Biological Sciences

Corresponding Author:rimadrid@ucm.es

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Abstract

Airbone viral pathogens can rapidly spread and become a global menace, including both human and animal viruses which can trigger important socioeconomical and health effects. Therefore, to prevent and contain potential epidemic outbreaks, accurate, fast, and affordable diagnostic point-of-care tests (POCT) are required. As a proof of concept, we have developed a molecular detection system based in Loop-mediated isothermal AMPlification technique (LAMP) for two different RNA airborne viruses: the well-known human SARS-CoV-2, and the avian metapneumovirus (aMPV), the aetiologic agent of a communicable disease infecting mainly turkeys and chickens. To obtain a POC diagnostic system, we coupled the LAMP technique to DNA-functionalized gold nanoparticles detection. Validation of this system was carried out in 140 pharyngeal swabs from COVID-19 symptomatic and asymptomatic patients, and in 50 different samples (pharyngeal swabs and tracheal tissue samples) collected from aMPV infected chickens and turkeys. The system allows viral detection in about 60 minutes by the naked eye with 100% specificity, and 97.22% and 87.88% sensitivity for SARS-CoV-2 and aMPV, respectively. In summary, this novel detection system based on the coupling of RT-LAMP to DNA-nanoprobes allows suitable virus testing in the field, with accurate levels very close to conventional qRT-PCR based diagnosis.