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

Phylogenetic analysis has demonstrated that the etiologic agent of the 2020 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 the Word Health Organ...

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Bibliographic Details
Published in:Scientific reports 2021-01, Vol.11 (1), p.2224-2224, Article 2224
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: Article
Language:English
Subjects:
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Complementarity
COVID-19
COVID-19 - diagnosis
COVID-19 Nucleic Acid Testing - methods
Diagnosis
Diagnostic tests
DNA, Single-Stranded
Enterovirus
Genome, Viral
Health promotion
Homology
Humanities and Social Sciences
Humans
multidisciplinary
Organic Chemicals
Pandemics
Phylogeny
Polymerase Chain Reaction
Public health
Real-Time Polymerase Chain Reaction - methods
Respiratory tract
Rhinovirus
SARS-CoV-2
Science
Science (multidisciplinary)
Sensitivity analysis
Sensitivity and Specificity
Severe acute respiratory syndrome coronavirus 2
Temperature
Viral Load
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Summary:Phylogenetic analysis has demonstrated that the etiologic agent of the 2020 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 the Word Health Organization (WHO) is RT-PCR. To detect low viral loads and perform large-scale screening, a low-cost diagnostic test is necessary. Here, we developed a cost-effective test capable of detecting SARS-CoV-2. We validated an auxiliary protocol for molecular diagnosis with the SYBR Green RT-PCR methodology to successfully screen negative cases of SARS-CoV-2. Our results revealed a set of primers with high specificity and no homology with other viruses from the Coronovideae family or human respiratory tract pathogenic viruses, presenting with complementarity only for rhinoviruses/enteroviruses and Legionella spp. Optimization of the annealing temperature and polymerization time led to a 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 a large scale to soften panic and economic burden through guidance for isolation strategies.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-021-81245-0