Development of a Fast and Affordable Diagnostic System for RNA Viruses
Using Loop-Mediated Isothermal Amplification and DNA-Nanoprobe Detection
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.