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Title:Robust saliva-based RNA extraction-free one-step nucleic acid amplification test for mass SARS-CoV-2 monitoring
Authors:ID Rajh, Eva (Author)
ID Šket, Tina (Author)
ID Praznik, Arne (Author)
ID Sušjan, Petra (Author)
ID Šmid, Alenka (Author)
ID Urbančič, Dunja (Author)
ID Mlinarič-Raščan, Irena (Author)
ID Kogovšek, Polona (Author)
ID Demšar, Tina (Author)
ID Milavec, Mojca (Author)
ID Prosenc, Katarina (Author)
ID Jensterle, Žiga (Author)
ID Zidarn, Mihaela, Klinika Golnik, Medicinska fakulteta UL (Author)
ID Tomič, Viktorija, Klinika Golnik (Author)
ID Turel, Gabriele (Author)
ID Lejko-Zupanc, Tatjana (Author)
ID Jerala, Roman (Author)
ID Benčina, Mojca (Author)
Files:URL URL - Presentation file, visit
Typology:1.01 - Original Scientific Article
Organization:Logo UKPBAG - University Clinic of Respiratory and Allergic Diseases Golnik
Abstract: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.
Keywords:SARS-CoV-2, COVID-19, COVID-19 serological testing, real-time polymerase chain reaction, saliva, oral cavity swab, passive drool, pooling
Publication status in journal:Published
Article version:Publisher's version of article
Place of publishing:Švica
Year of publishing:2021
Number of pages:str. 1-19
Numbering:Vol. 26, iss. 21
PID:20.500.12556/DiRROS-14590 New window
ISSN on article:1420-3049
DOI:10.3390/molecules26216617 New window
COBISS.SI-ID:83744003 New window
Copyright:© by the authors
Note:Nasl. z nasl. zaslona; Soavtorji: 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; Opis vira z dne 5. 11. 2021; Eva Rajh in Tina Šket sta enakovredni prvi avtorici; Št. članka: 6617;
Publication date in DiRROS:09.11.2021
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Shortened title:Molecules
COBISS.SI-ID:18462981 New window


License:CC BY 4.0, Creative Commons Attribution 4.0 International
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:31.10.2021

Secondary language

Keywords:SARS-CoV-2, covid-19, serološko testiranje za COVID-19, verižna reakcija s polimerazo v realnem času, slina, bris ustne sluznice, združeni vzorci