A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva

Detalhes bibliográficos
Autor(a) principal: Da Silva, Sofia M.
Data de Publicação: 2024
Outros Autores: Amaral, Catarina, Malta-Luís, Cláudia, Grilo, Diana, Duarte, Americo G., Morais, Inês, Afonso, Gonçalo, Faria, Nuno, Antunes, Wilson, Gomes, Inês, Sá-Leão, Raquel, Miragaia, Maria, Serrano, Mónica, Pimentel, Catarina
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Texto Completo: http://hdl.handle.net/10362/172778
Resumo: Funding Information: This work was supported by (i) Project LISBOA-01-0145-FEDER-007660 (\u2018Microbiologia Molecular, Estrutural e Celular\u2019) funded by FEDER \u2018Fundo Europeu de Desenvolvimento Regional\u2019 funds through COMPETE2020\u2014\u2018Programa Operacional Competitividade e Internacionaliza\u00E7\u00E3o\u2019 (POCI), (ii) \u2018Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia\u2019 (FCT) through the project DETECT Ref 433_613549914 (20/7/153), under the scope of the 2nd edition of the programme RESEARCH4COVID19, (iii) the project \u2018STOP-COVID\u2014Strategies to prevent COVID-19 by early detection of asymptomatic carriers at increased risk: epidemiological studies and validation of a rapid in-house diagnostic test\u2019, Ref 072559, funded by FEDER from \u2018Programa Operacional Regional Lisboa\u2019 and (iv) the Municipality of Oeiras. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Funding Information: This work was supported by (i) Project LISBOA-01-0145-FEDER- 007660 ( Microbiologia Molecular, Estrutural e Celular ) funded by FEDER Fundo Europeu de Desenvolvimento Regional funds through COMPETE2020\u2014 Programa Operacional Competitividade e Internacionaliza\u00E7ao ~ (POCI), (ii) Funda\u00E7ao ~ para a Ciencia ^ e a Tecnologia (FCT) through the project DETECT Ref 433_613549914 (20/7/153), under the scope of the 2nd edition of the programme RESEARCH4COVID19, (iii) the project STOP-COVID\u2014Strategies to prevent COVID-19 by early detection of asymptomatic carriers at increased risk: epidemiological studies and validation of a rapid in-house diagnostic test , Ref 072559, funded by FEDER from Programa Operacional Regional Lisboa and (iv) the Municipality of Oeiras. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Publisher Copyright: © 2024 The Author(s). Published by Oxford University Press.
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spelling A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed salivaCOVID-19diagnosticsRT-PCR SYBR Greensaliva testBiochemistry, Genetics and Molecular Biology(all)Agricultural and Biological Sciences(all)SDG 3 - Good Health and Well-beingFunding Information: This work was supported by (i) Project LISBOA-01-0145-FEDER-007660 (\u2018Microbiologia Molecular, Estrutural e Celular\u2019) funded by FEDER \u2018Fundo Europeu de Desenvolvimento Regional\u2019 funds through COMPETE2020\u2014\u2018Programa Operacional Competitividade e Internacionaliza\u00E7\u00E3o\u2019 (POCI), (ii) \u2018Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia\u2019 (FCT) through the project DETECT Ref 433_613549914 (20/7/153), under the scope of the 2nd edition of the programme RESEARCH4COVID19, (iii) the project \u2018STOP-COVID\u2014Strategies to prevent COVID-19 by early detection of asymptomatic carriers at increased risk: epidemiological studies and validation of a rapid in-house diagnostic test\u2019, Ref 072559, funded by FEDER from \u2018Programa Operacional Regional Lisboa\u2019 and (iv) the Municipality of Oeiras. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Funding Information: This work was supported by (i) Project LISBOA-01-0145-FEDER- 007660 ( Microbiologia Molecular, Estrutural e Celular ) funded by FEDER Fundo Europeu de Desenvolvimento Regional funds through COMPETE2020\u2014 Programa Operacional Competitividade e Internacionaliza\u00E7ao ~ (POCI), (ii) Funda\u00E7ao ~ para a Ciencia ^ e a Tecnologia (FCT) through the project DETECT Ref 433_613549914 (20/7/153), under the scope of the 2nd edition of the programme RESEARCH4COVID19, (iii) the project STOP-COVID\u2014Strategies to prevent COVID-19 by early detection of asymptomatic carriers at increased risk: epidemiological studies and validation of a rapid in-house diagnostic test , Ref 072559, funded by FEDER from Programa Operacional Regional Lisboa and (iv) the Municipality of Oeiras. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Publisher Copyright: © 2024 The Author(s). Published by Oxford University Press.The gold standard for coronavirus disease 2019 diagnostic testing relies on RNA extraction from naso/oropharyngeal swab followed by amplification through reverse transcription-polymerase chain reaction (RT-PCR) with fluorogenic probes. While the test is extremely sensitive and specific, its high cost and the potential discomfort associated with specimen collection made it suboptimal for public health screening purposes. In this study, we developed an equally reliable, but cheaper and less invasive alternative test based on a one-step RT-PCR with the DNA-intercalating dye SYBR Green, which enables the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) directly from saliva samples or RNA isolated from nasopharyngeal (NP) swabs. Importantly, we found that this type of testing can be fine-tuned to discriminate SARS-CoV-2 variants of concern. The saliva RT-PCR SYBR Green test was successfully used in a mass-screening initiative targeting nearly 4500 asymptomatic children under the age of 12. Testing was performed at a reasonable cost, and in some cases, the saliva test outperformed NP rapid antigen tests in identifying infected children. Whole genome sequencing revealed that the antigen testing failure could not be attributed to a specific lineage of SARS-CoV-2. Overall, this work strongly supports the view that RT-PCR saliva tests based on DNA-intercalating dyes represent a powerful strategy for community screening of SARS-CoV-2. The tests can be easily applied to other infectious agents and, therefore, constitute a powerful resource for an effective response to future pandemics.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNDa Silva, Sofia M.Amaral, CatarinaMalta-Luís, CláudiaGrilo, DianaDuarte, Americo G.Morais, InêsAfonso, GonçaloFaria, NunoAntunes, WilsonGomes, InêsSá-Leão, RaquelMiragaia, MariaSerrano, MónicaPimentel, Catarina2024-09-30T22:35:37Z2024-05-222024-05-22T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/172778eng2396-8923PURE: 100336126https://doi.org/10.1093/biomethods/bpae035info:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2025-03-31T01:53:43Zoai:run.unl.pt:10362/172778Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-28T18:55:22.491230Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
title A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
spellingShingle A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
Da Silva, Sofia M.
COVID-19
diagnostics
RT-PCR SYBR Green
saliva test
Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)
SDG 3 - Good Health and Well-being
title_short A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
title_full A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
title_fullStr A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
title_full_unstemmed A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
title_sort A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
author Da Silva, Sofia M.
author_facet Da Silva, Sofia M.
Amaral, Catarina
Malta-Luís, Cláudia
Grilo, Diana
Duarte, Americo G.
Morais, Inês
Afonso, Gonçalo
Faria, Nuno
Antunes, Wilson
Gomes, Inês
Sá-Leão, Raquel
Miragaia, Maria
Serrano, Mónica
Pimentel, Catarina
author_role author
author2 Amaral, Catarina
Malta-Luís, Cláudia
Grilo, Diana
Duarte, Americo G.
Morais, Inês
Afonso, Gonçalo
Faria, Nuno
Antunes, Wilson
Gomes, Inês
Sá-Leão, Raquel
Miragaia, Maria
Serrano, Mónica
Pimentel, Catarina
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
RUN
dc.contributor.author.fl_str_mv Da Silva, Sofia M.
Amaral, Catarina
Malta-Luís, Cláudia
Grilo, Diana
Duarte, Americo G.
Morais, Inês
Afonso, Gonçalo
Faria, Nuno
Antunes, Wilson
Gomes, Inês
Sá-Leão, Raquel
Miragaia, Maria
Serrano, Mónica
Pimentel, Catarina
dc.subject.por.fl_str_mv COVID-19
diagnostics
RT-PCR SYBR Green
saliva test
Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)
SDG 3 - Good Health and Well-being
topic COVID-19
diagnostics
RT-PCR SYBR Green
saliva test
Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)
SDG 3 - Good Health and Well-being
description Funding Information: This work was supported by (i) Project LISBOA-01-0145-FEDER-007660 (\u2018Microbiologia Molecular, Estrutural e Celular\u2019) funded by FEDER \u2018Fundo Europeu de Desenvolvimento Regional\u2019 funds through COMPETE2020\u2014\u2018Programa Operacional Competitividade e Internacionaliza\u00E7\u00E3o\u2019 (POCI), (ii) \u2018Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia\u2019 (FCT) through the project DETECT Ref 433_613549914 (20/7/153), under the scope of the 2nd edition of the programme RESEARCH4COVID19, (iii) the project \u2018STOP-COVID\u2014Strategies to prevent COVID-19 by early detection of asymptomatic carriers at increased risk: epidemiological studies and validation of a rapid in-house diagnostic test\u2019, Ref 072559, funded by FEDER from \u2018Programa Operacional Regional Lisboa\u2019 and (iv) the Municipality of Oeiras. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Funding Information: This work was supported by (i) Project LISBOA-01-0145-FEDER- 007660 ( Microbiologia Molecular, Estrutural e Celular ) funded by FEDER Fundo Europeu de Desenvolvimento Regional funds through COMPETE2020\u2014 Programa Operacional Competitividade e Internacionaliza\u00E7ao ~ (POCI), (ii) Funda\u00E7ao ~ para a Ciencia ^ e a Tecnologia (FCT) through the project DETECT Ref 433_613549914 (20/7/153), under the scope of the 2nd edition of the programme RESEARCH4COVID19, (iii) the project STOP-COVID\u2014Strategies to prevent COVID-19 by early detection of asymptomatic carriers at increased risk: epidemiological studies and validation of a rapid in-house diagnostic test , Ref 072559, funded by FEDER from Programa Operacional Regional Lisboa and (iv) the Municipality of Oeiras. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Publisher Copyright: © 2024 The Author(s). Published by Oxford University Press.
publishDate 2024
dc.date.none.fl_str_mv 2024-09-30T22:35:37Z
2024-05-22
2024-05-22T00:00:00Z
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PURE: 100336126
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