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Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring

Early diagnosis with rapid detection of the virus plays a key role in preventing the spread of infection and in treating patients effectively. In order to address the need for a straightforward detection of SARS-CoV-2 infection and assessment of viral spread, we developed rapid, sensitive, extractio...

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Autores principales: Rajh, Eva, Šket, Tina, Praznik, Arne, Sušjan, Petra, Šmid, Alenka, Urbančič, Dunja, Mlinarič-Raščan, Irena, Kogovšek, Polona, Demšar, Tina, Milavec, Mojca, Prosenc Trilar, Katarina, Jensterle, Žiga, Zidarn, Mihaela, Tomič, Viktorija, Turel, Gabriele, Lejko-Zupanc, Tatjana, Jerala, Roman, Benčina, Mojca
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8588466/
https://www.ncbi.nlm.nih.gov/pubmed/34771026
http://dx.doi.org/10.3390/molecules26216617
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author Rajh, Eva
Šket, Tina
Praznik, Arne
Sušjan, Petra
Šmid, Alenka
Urbančič, Dunja
Mlinarič-Raščan, Irena
Kogovšek, Polona
Demšar, Tina
Milavec, Mojca
Prosenc Trilar, Katarina
Jensterle, Žiga
Zidarn, Mihaela
Tomič, Viktorija
Turel, Gabriele
Lejko-Zupanc, Tatjana
Jerala, Roman
Benčina, Mojca
author_facet Rajh, Eva
Šket, Tina
Praznik, Arne
Sušjan, Petra
Šmid, Alenka
Urbančič, Dunja
Mlinarič-Raščan, Irena
Kogovšek, Polona
Demšar, Tina
Milavec, Mojca
Prosenc Trilar, Katarina
Jensterle, Žiga
Zidarn, Mihaela
Tomič, Viktorija
Turel, Gabriele
Lejko-Zupanc, Tatjana
Jerala, Roman
Benčina, Mojca
author_sort Rajh, Eva
collection PubMed
description Early diagnosis with rapid detection of the virus plays a key role in preventing the spread of infection and in treating patients effectively. In order to address the need for a straightforward detection of SARS-CoV-2 infection and assessment of viral spread, we developed rapid, sensitive, extraction-free one-step reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) tests for detecting SARS-CoV-2 in saliva. We analyzed over 700 matched pairs of saliva and nasopharyngeal swab (NSB) specimens from asymptomatic and symptomatic individuals. Saliva, as either an oral cavity swab or passive drool, was collected in an RNA stabilization buffer. The stabilized saliva specimens were heat-treated and directly analyzed without RNA extraction. The diagnostic sensitivity of saliva-based RT-qPCR was at least 95% in individuals with subclinical infection and outperformed RT-LAMP, which had at least 70% sensitivity when compared to NSBs analyzed with a clinical RT-qPCR test. The diagnostic sensitivity for passive drool saliva was higher than that of oral cavity swab specimens (95% and 87%, respectively). A rapid, sensitive one-step extraction-free RT-qPCR test for detecting SARS-CoV-2 in passive drool saliva is operationally simple and can be easily implemented using existing testing sites, thus allowing high-throughput, rapid, and repeated testing of large populations. Furthermore, saliva testing is adequate to detect individuals in an asymptomatic screening program and can help improve voluntary screening compliance for those individuals averse to various forms of nasal collections.
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spelling pubmed-85884662021-11-13 Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring Rajh, Eva Šket, Tina Praznik, Arne Sušjan, Petra Šmid, Alenka Urbančič, Dunja Mlinarič-Raščan, Irena Kogovšek, Polona Demšar, Tina Milavec, Mojca Prosenc Trilar, Katarina Jensterle, Žiga Zidarn, Mihaela Tomič, Viktorija Turel, Gabriele Lejko-Zupanc, Tatjana Jerala, Roman Benčina, Mojca Molecules Article Early diagnosis with rapid detection of the virus plays a key role in preventing the spread of infection and in treating patients effectively. In order to address the need for a straightforward detection of SARS-CoV-2 infection and assessment of viral spread, we developed rapid, sensitive, extraction-free one-step reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) tests for detecting SARS-CoV-2 in saliva. We analyzed over 700 matched pairs of saliva and nasopharyngeal swab (NSB) specimens from asymptomatic and symptomatic individuals. Saliva, as either an oral cavity swab or passive drool, was collected in an RNA stabilization buffer. The stabilized saliva specimens were heat-treated and directly analyzed without RNA extraction. The diagnostic sensitivity of saliva-based RT-qPCR was at least 95% in individuals with subclinical infection and outperformed RT-LAMP, which had at least 70% sensitivity when compared to NSBs analyzed with a clinical RT-qPCR test. The diagnostic sensitivity for passive drool saliva was higher than that of oral cavity swab specimens (95% and 87%, respectively). A rapid, sensitive one-step extraction-free RT-qPCR test for detecting SARS-CoV-2 in passive drool saliva is operationally simple and can be easily implemented using existing testing sites, thus allowing high-throughput, rapid, and repeated testing of large populations. Furthermore, saliva testing is adequate to detect individuals in an asymptomatic screening program and can help improve voluntary screening compliance for those individuals averse to various forms of nasal collections. MDPI 2021-10-31 /pmc/articles/PMC8588466/ /pubmed/34771026 http://dx.doi.org/10.3390/molecules26216617 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rajh, Eva
Šket, Tina
Praznik, Arne
Sušjan, Petra
Šmid, Alenka
Urbančič, Dunja
Mlinarič-Raščan, Irena
Kogovšek, Polona
Demšar, Tina
Milavec, Mojca
Prosenc Trilar, Katarina
Jensterle, Žiga
Zidarn, Mihaela
Tomič, Viktorija
Turel, Gabriele
Lejko-Zupanc, Tatjana
Jerala, Roman
Benčina, Mojca
Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring
title Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring
title_full Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring
title_fullStr Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring
title_full_unstemmed Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring
title_short Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring
title_sort robust saliva-based rna extraction-free one-step nucleic acid amplification test for mass sars-cov-2 monitoring
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8588466/
https://www.ncbi.nlm.nih.gov/pubmed/34771026
http://dx.doi.org/10.3390/molecules26216617
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