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:	Eva Rajh, Tina Šket, Arne Praznik, Petra Sušjan, Alenka Šmid, Dunja Urbančič, Irena Mlinarič-Raščan, Polona Kogovšek, Tina Demšar, Mojca Milavec, Katarina Prosenc Trilar, Žiga Jensterle, Mihaela Zidarn, Viktorija Tomič, Gabriele Turel, Tatjana Lejko-Zupanc, Roman Jerala, Mojca Benčina
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	saliva COVID-19 SARS-CoV-2 LAMP RT-qPCR passive drool Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/42c6d2b103974f368a8c657759dd6512
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spelling	oai:doaj.org-article:42c6d2b103974f368a8c657759dd65122021-11-11T18:35:22ZRobust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring10.3390/molecules262166171420-3049https://doaj.org/article/42c6d2b103974f368a8c657759dd65122021-10-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/21/6617https://doaj.org/toc/1420-3049Early 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.Eva RajhTina ŠketArne PraznikPetra SušjanAlenka ŠmidDunja UrbančičIrena Mlinarič-RaščanPolona KogovšekTina DemšarMojca MilavecKatarina Prosenc TrilarŽiga JensterleMihaela ZidarnViktorija TomičGabriele TurelTatjana Lejko-ZupancRoman JeralaMojca BenčinaMDPI AGarticlesalivaCOVID-19SARS-CoV-2LAMPRT-qPCRpassive droolOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6617, p 6617 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	saliva COVID-19 SARS-CoV-2 LAMP RT-qPCR passive drool Organic chemistry QD241-441
spellingShingle	saliva COVID-19 SARS-CoV-2 LAMP RT-qPCR passive drool Organic chemistry QD241-441 Eva Rajh Tina Šket Arne Praznik Petra Sušjan Alenka Šmid Dunja Urbančič Irena Mlinarič-Raščan Polona Kogovšek Tina Demšar Mojca Milavec Katarina Prosenc Trilar Žiga Jensterle Mihaela Zidarn Viktorija Tomič Gabriele Turel Tatjana Lejko-Zupanc Roman Jerala Mojca Benčina Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring
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.
format	article
author	Eva Rajh Tina Šket Arne Praznik Petra Sušjan Alenka Šmid Dunja Urbančič Irena Mlinarič-Raščan Polona Kogovšek Tina Demšar Mojca Milavec Katarina Prosenc Trilar Žiga Jensterle Mihaela Zidarn Viktorija Tomič Gabriele Turel Tatjana Lejko-Zupanc Roman Jerala Mojca Benčina
author_facet	Eva Rajh Tina Šket Arne Praznik Petra Sušjan Alenka Šmid Dunja Urbančič Irena Mlinarič-Raščan Polona Kogovšek Tina Demšar Mojca Milavec Katarina Prosenc Trilar Žiga Jensterle Mihaela Zidarn Viktorija Tomič Gabriele Turel Tatjana Lejko-Zupanc Roman Jerala Mojca Benčina
author_sort	Eva Rajh
title	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_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_sort	robust saliva-based rna extraction-free one-step nucleic acid amplification test for mass sars-cov-2 monitoring
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/42c6d2b103974f368a8c657759dd6512
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Robust Saliva-Based RNA Extraction-Free One-Step Nucleic Acid Amplification Test for Mass SARS-CoV-2 Monitoring

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