Cargando…
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...
Autores principales: | , , , , , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1784598467540156416 |
---|---|
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. |
format | Online Article Text |
id | pubmed-8588466 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT rajheva robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT skettina robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT praznikarne robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT susjanpetra robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT smidalenka robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT urbancicdunja robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT mlinaricrascanirena robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT kogovsekpolona robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT demsartina robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT milavecmojca robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT prosenctrilarkatarina robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT jensterleziga robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT zidarnmihaela robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT tomicviktorija robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT turelgabriele robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT lejkozupanctatjana robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT jeralaroman robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring AT bencinamojca robustsalivabasedrnaextractionfreeonestepnucleicacidamplificationtestformasssarscov2monitoring |