Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection

The fast spread of SARS-CoV-2 has led to a global pandemic, calling for fast and accurate assays to allow infection diagnosis and prevention of transmission. We aimed to develop a molecular beacon (MB)-based detection assay for SARS-CoV-2, designed to detect the ORF1ab and S genes, proposing a two-s...

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Autores principales:	Josué Carvalho, Jéssica Lopes-Nunes, Joana Figueiredo, Tiago Santos, André Miranda, Micaela Riscado, Fani Sousa, Ana Paula Duarte, Sílvia Socorro, Cândida Teixeira Tomaz, Mafalda Felgueiras, Rui Teixeira, Conceição Faria, Carla Cruz
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	RNA viruses SARS-CoV-2 COVID-19 fluorescence detection diagnosis Chemical technology TP1-1185
Acceso en línea:	https://doaj.org/article/46073fa94116449d9743c9201aa73dd0
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spelling	oai:doaj.org-article:46073fa94116449d9743c9201aa73dd02021-11-11T19:03:09ZMolecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection10.3390/s212170151424-8220https://doaj.org/article/46073fa94116449d9743c9201aa73dd02021-10-01T00:00:00Zhttps://www.mdpi.com/1424-8220/21/21/7015https://doaj.org/toc/1424-8220The fast spread of SARS-CoV-2 has led to a global pandemic, calling for fast and accurate assays to allow infection diagnosis and prevention of transmission. We aimed to develop a molecular beacon (MB)-based detection assay for SARS-CoV-2, designed to detect the ORF1ab and S genes, proposing a two-stage COVID-19 testing strategy. The novelty of this work lies in the design and optimization of two MBs for detection of SARS-CoV-2, namely, concentration, fluorescence plateaus of hybridization, reaction temperature and real-time results. We also identify putative G-quadruplex (G4) regions in the genome of SARS-CoV-2. A total of 458 nasopharyngeal and throat swab samples (426 positive and 32 negative) were tested with the MB assay and the fluorescence levels compared with the cycle threshold (Ct) values obtained from a commercial RT-PCR test in terms of test duration, sensitivity, and specificity. Our results show that the samples with higher fluorescence levels correspond to those with low Ct values, suggesting a correlation between viral load and increased MB fluorescence. The proposed assay represents a fast (total duration of 2 h 20 min including amplification and fluorescence reading stages) and simple way of detecting SARS-CoV-2 in clinical samples from the upper respiratory tract.Josué CarvalhoJéssica Lopes-NunesJoana FigueiredoTiago SantosAndré MirandaMicaela RiscadoFani SousaAna Paula DuarteSílvia SocorroCândida Teixeira TomazMafalda FelgueirasRui TeixeiraConceição FariaCarla CruzMDPI AGarticleRNA virusesSARS-CoV-2COVID-19fluorescence detectiondiagnosisChemical technologyTP1-1185ENSensors, Vol 21, Iss 7015, p 7015 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	RNA viruses SARS-CoV-2 COVID-19 fluorescence detection diagnosis Chemical technology TP1-1185
spellingShingle	RNA viruses SARS-CoV-2 COVID-19 fluorescence detection diagnosis Chemical technology TP1-1185 Josué Carvalho Jéssica Lopes-Nunes Joana Figueiredo Tiago Santos André Miranda Micaela Riscado Fani Sousa Ana Paula Duarte Sílvia Socorro Cândida Teixeira Tomaz Mafalda Felgueiras Rui Teixeira Conceição Faria Carla Cruz Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection
description	The fast spread of SARS-CoV-2 has led to a global pandemic, calling for fast and accurate assays to allow infection diagnosis and prevention of transmission. We aimed to develop a molecular beacon (MB)-based detection assay for SARS-CoV-2, designed to detect the ORF1ab and S genes, proposing a two-stage COVID-19 testing strategy. The novelty of this work lies in the design and optimization of two MBs for detection of SARS-CoV-2, namely, concentration, fluorescence plateaus of hybridization, reaction temperature and real-time results. We also identify putative G-quadruplex (G4) regions in the genome of SARS-CoV-2. A total of 458 nasopharyngeal and throat swab samples (426 positive and 32 negative) were tested with the MB assay and the fluorescence levels compared with the cycle threshold (Ct) values obtained from a commercial RT-PCR test in terms of test duration, sensitivity, and specificity. Our results show that the samples with higher fluorescence levels correspond to those with low Ct values, suggesting a correlation between viral load and increased MB fluorescence. The proposed assay represents a fast (total duration of 2 h 20 min including amplification and fluorescence reading stages) and simple way of detecting SARS-CoV-2 in clinical samples from the upper respiratory tract.
format	article
author	Josué Carvalho Jéssica Lopes-Nunes Joana Figueiredo Tiago Santos André Miranda Micaela Riscado Fani Sousa Ana Paula Duarte Sílvia Socorro Cândida Teixeira Tomaz Mafalda Felgueiras Rui Teixeira Conceição Faria Carla Cruz
author_facet	Josué Carvalho Jéssica Lopes-Nunes Joana Figueiredo Tiago Santos André Miranda Micaela Riscado Fani Sousa Ana Paula Duarte Sílvia Socorro Cândida Teixeira Tomaz Mafalda Felgueiras Rui Teixeira Conceição Faria Carla Cruz
author_sort	Josué Carvalho
title	Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection
title_short	Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection
title_full	Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection
title_fullStr	Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection
title_full_unstemmed	Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection
title_sort	molecular beacon assay development for severe acute respiratory syndrome coronavirus 2 detection
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/46073fa94116449d9743c9201aa73dd0
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Molecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detection

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