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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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Autores principales: Marinowic, D. R., Zanirati, G., Rodrigues, F. V. F., Grahl, M. V. C., Alcará, A. M., Machado, D. C., Da Costa, J. C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7838253/
https://www.ncbi.nlm.nih.gov/pubmed/33500453
http://dx.doi.org/10.1038/s41598-021-81245-0
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author Marinowic, D. R.
Zanirati, G.
Rodrigues, F. V. F.
Grahl, M. V. C.
Alcará, A. M.
Machado, D. C.
Da Costa, J. C.
author_facet Marinowic, D. R.
Zanirati, G.
Rodrigues, F. V. F.
Grahl, M. V. C.
Alcará, A. M.
Machado, D. C.
Da Costa, J. C.
author_sort Marinowic, D. R.
collection PubMed
description 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.
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spelling pubmed-78382532021-01-27 A new SYBR Green real-time PCR to detect SARS-CoV-2 Marinowic, D. R. Zanirati, G. Rodrigues, F. V. F. Grahl, M. V. C. Alcará, A. M. Machado, D. C. Da Costa, J. C. Sci Rep Article 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. Nature Publishing Group UK 2021-01-26 /pmc/articles/PMC7838253/ /pubmed/33500453 http://dx.doi.org/10.1038/s41598-021-81245-0 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Marinowic, D. R.
Zanirati, G.
Rodrigues, F. V. F.
Grahl, M. V. C.
Alcará, A. M.
Machado, D. C.
Da Costa, J. C.
A new SYBR Green real-time PCR to detect SARS-CoV-2
title A new SYBR Green real-time PCR to detect SARS-CoV-2
title_full A new SYBR Green real-time PCR to detect SARS-CoV-2
title_fullStr A new SYBR Green real-time PCR to detect SARS-CoV-2
title_full_unstemmed A new SYBR Green real-time PCR to detect SARS-CoV-2
title_short A new SYBR Green real-time PCR to detect SARS-CoV-2
title_sort new sybr green real-time pcr to detect sars-cov-2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7838253/
https://www.ncbi.nlm.nih.gov/pubmed/33500453
http://dx.doi.org/10.1038/s41598-021-81245-0
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