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

2021-10Journal article Open access
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dc.contributor.authorCarvalho, JosuƩ
dc.contributor.authorNunes, J. Lopes
dc.contributor.authorFigueiredo, Joana
dc.contributor.authorSantos, Tiago
dc.contributor.authorMiranda, AndrƩ
dc.contributor.authorRiscado, Micaela
dc.contributor.authorSousa, Fani
dc.contributor.authorDuarte, A. P.
dc.contributor.authorSocorro, SĆ­lvia
dc.contributor.authorTomaz, CĆ¢ndida
dc.contributor.authorFelgueiras, Mafalda
dc.contributor.authorTeixeira, Rui
dc.contributor.authorFaria, ConceiĆ§Ć£o
dc.contributor.authorCruz, Carla
dc.date.accessioned2022-01-20T15:35:49Z
dc.date.available2022-01-20T15:35:49Z
dc.date.issued2021-10
dc.description.abstractThe 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.pt_PT
dc.description.sponsorshipThis work was supported by project No. 149 ā€œTrack and Trace COVID-19ā€ funded in first edition ā€œRESEARCH 4 COVID-19ā€ by FundaĆ§Ć£o para a CiĆŖncia e a Tecnologia (FCT) in collaboration with AgĆŖncia de InvestigaĆ§Ć£o ClĆ­nica e InovaĆ§Ć£o BiomĆ©dica (AICIB)pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.3390/s21217015pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/11930
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.relationHealth Sciences Research Centre
dc.relationG-quadruplex aptamers for cervical cancer therapy
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectRNA virusespt_PT
dc.subjectSARS-CoV-2pt_PT
dc.subjectCOVID-19pt_PT
dc.subjectFluorescence detectionpt_PT
dc.subjectDiagnosispt_PT
dc.titleMolecular Beacon Assay Development for Severe Acute Respiratory Syndrome Coronavirus 2 Detectionpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleHealth Sciences Research Centre
oaire.awardTitleG-quadruplex aptamers for cervical cancer therapy
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00709%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F122953%2F2016/PT
oaire.citation.issue21pt_PT
oaire.citation.startPage7015pt_PT
oaire.citation.titleSensorspt_PT
oaire.citation.volume21pt_PT
oaire.fundingStream6817 - DCRRNI ID
person.familyNamede Oliveira e Carvalho
person.familyNameNunes
person.familyNameAfonso dos Santos
person.familyNameRodrigues Miranda
person.familyNameRibeiro Riscado
person.familyNameSousa
person.familyNameDuarte
person.familyNameSocorro
person.familyNameTomaz
person.familyNameCruz
person.givenNameJosuƩ Leandro
person.givenNameJĆ©ssica
person.givenNameTiago AndrƩ
person.givenNameAndrƩ Filipe
person.givenNameMicaela Sofia
person.givenNameFani
person.givenNameAna Paula
person.givenNameSĆ­lvia Cristina da Cruz Marques
person.givenNameCĆ¢ndida
person.givenNameCarla
person.identifier1255684
person.identifierhttps://scholar.google.pt/citations?user=PA3L_h8AAAAJ&hl=pt-PT
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project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundaĆ§Ć£o para a CiĆŖncia e a Tecnologia
project.funder.nameFundaĆ§Ć£o para a CiĆŖncia e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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