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

Abstract 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 Hea...

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Autores principales:	D. R. Marinowic, G. Zanirati, F. V. F. Rodrigues, M. V. C. Grahl, A. M. Alcará, D. C. Machado, J. C. Da Costa
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7d5be5345bc64a98a88b288631832959
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spelling	oai:doaj.org-article:7d5be5345bc64a98a88b2886318329592021-12-02T13:57:26ZA new SYBR Green real-time PCR to detect SARS-CoV-210.1038/s41598-021-81245-02045-2322https://doaj.org/article/7d5be5345bc64a98a88b2886318329592021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81245-0https://doaj.org/toc/2045-2322Abstract 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.D. R. MarinowicG. ZaniratiF. V. F. RodriguesM. V. C. GrahlA. M. AlcaráD. C. MachadoJ. C. Da CostaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q D. R. Marinowic G. Zanirati F. V. F. Rodrigues M. V. C. Grahl A. M. Alcará D. C. Machado J. C. Da Costa A new SYBR Green real-time PCR to detect SARS-CoV-2
description	Abstract 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.
format	article
author	D. R. Marinowic G. Zanirati F. V. F. Rodrigues M. V. C. Grahl A. M. Alcará D. C. Machado J. C. Da Costa
author_facet	D. R. Marinowic G. Zanirati F. V. F. Rodrigues M. V. C. Grahl A. M. Alcará D. C. Machado J. C. Da Costa
author_sort	D. R. Marinowic
title	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_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_sort	new sybr green real-time pcr to detect sars-cov-2
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/7d5be5345bc64a98a88b288631832959
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A new SYBR Green real-time PCR to detect SARS-CoV-2

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