A new Syber Green real time PCR to detect SARS-CoV-2

• Published in: Nature Portfolio 2020
• Main Authors: Marinowic, D R, Zanirati, G, Rodrigues, F.V.F., Grahl, M.V.C., Alcará, A M, Machado, D C, Da Costa, J.C.
• Format: Text Resource
• Language: English
• Subjects: Diagnostic tests; Severe acute respiratory syndrome coronavirus 2
• DOI: 10.21203/rs.3.rs-28188/v1

Phylogenetic analyses demonstrated that etiologic agent of pandemic outbreak is a betacoronavirus named SARS-CoV-2. For public health interventions, a diagnostic test with high sensitivity and specificity is required. The gold standard protocol for diagnosis by WHO is the RT-PCR. To detect low viral load and large-scale screening a low-cost diagnostic test becomes necessary. Here we develop a cost-effective test capable of to detect the new coronavirues. We validated an auxiliary protocol for molecular diagnosis with RT-PCR SYBR Green methodology to successfully screen negative cases of SARS-CoV-2. Our results demonstrated that a set of primers with high specificity, and no homology with other viruses from Coronovideae family or human respiratory tract pathogenic viruses. Optimization of annealing temperature and polymerization time led to an high specificity in the PCR products. We have developed a more affordable and swift methodology for negative SARS-CoV-2 screening. This methodology can be applied on large scale populational to soften panic and economic burden through guidance for isolation strategies.