A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2024 |
| Outros Autores: | , , , , , , , , , , , , |
| 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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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. |
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2024 |
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2024-09-30T22:35:37Z 2024-05-22 2024-05-22T00:00:00Z |
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2396-8923 PURE: 100336126 https://doi.org/10.1093/biomethods/bpae035 |
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