V.M. Lugo-Ramírez, R.G. Ramírez-Chavarría, E. Castillo-Villanueva
Universidad Nacional Autónoma de México,
Keywords: fluorescence quantification,influenza, LAMP, pathogen detection, portable device, SARS-CoV-2
Summary:Currently, influenza and coronavirus (SARS-CoV-2) infections present similar symptoms. For this reason, accurate and rapid diagnoses are needed to guide decision-making regarding hospital admission, specific antiviral treatments, and infection control measures. While polymerase chain reaction (PCR) remains the gold standard for molecular assays, its complexity makes its implementation difficult in devices that can be used outside laboratories and without trained personnel. Recent development of isothermal amplification techniques enables the integration of these techniques into microsystems and portable devices that allow in-situ tests to be carried out. In this work, we present the design of a device for the simultaneous detection of the Influenza A and SARS-CoV-2 viruses by the loop-mediated isothermal amplification (LAMP). Primers designed for each virus are marked with specific fluorophores (FAM and ROX). During the amplification process, fluorescence is quantified and data is transmitted to a computer using an application developed using LabVIEW. Through the response characterization of samples with known viral loads, we established a calibration curve that allows estimating the viral load in subsequently analyzed samples. The designed device has low fabrication costs and complexity, it has also high specificity and stands out for its versatility. Finally, we devise the device usage with the correct primer design to detect multiple pathogens in samples from various origins.