Desenvolvimento e construção de plataformas analíticas portáteis para detecção de RNA viral visando o diagnóstico de doenças infecciosas

Abstract

Viral respiratory infections represent a major threat to the health of the population around the world. In many cases, COVID-19 presents symptoms that can be confused with Influenza, a disease caused by the Influenza A virus. A simple, rapid, and selective assay, capable of identifying the etiological agent and differentiating the diseases, is essential to provide the correct management clinician to the patient. Here, we describe the development of a genomic assay for the selective capture of SARS-CoV-2 and Influenza A virus RNA in saliva samples using two portable analysis systems, consisting of a platform for electrochemical detection (PDE) and a platform with digital image detection (PID). The proposed methods showed excellent performance in the detection and quantification of viral RNA, with PDE presenting a limit of detection (LoD) and limit of quantification (LoQ) of 5.0 fmol L-1 and 8.6 fmol L-1 for SARS-CoV-2 and 1.0 fmol L-1 and 108.9 fmol L-1 for Influenza, respectively. PID was applied to detect the Influenza A virus and presented LoD and LoQ for 0.093 pmol L-1 and 0.310 pmol L-1, respectively. Genomagnetic assays were used to evaluate the presence of viruses in 36 saliva samples from volunteers and the results were in agreement with those obtained by reverse transcription polymerase chain reaction (RT-PCR), demonstrating the viability of the proposed methods for diagnosis. of COVID-19 and Influenza with point-of-care capability.