Robust saliva-based RNA extraction-free one-step nucleic acid amplification test for mass SARS-CoV-2 monitoring
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 Trilar, Katarina (Author), ID Jensterle, Žiga (Author), ID Zidarn, Mihaela (Author), ID Tomič, Viktorija (Author), ID Turel, Gabriele (Author), ID Lejko-Zupanc, Tatjana (Author), ID Jerala, Roman (Author), ID Benčina, Mojca (Author)

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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:oral cavity swab, passive drool, pooling, saliva, COVID-19, SARS-CoV-2, LAMP, RT-qPCR, COVID-19 serological testing, real-time polymerase chain reaction
Work type:Article
Typology:1.01 - Original Scientific Article
Organization:FFA - Faculty of Pharmacy
BF - Biotechnical Faculty
Publication status:Published
Publication version:Version of Record
Number of pages:19 str.
Numbering:Vol. 26, iss. 21, art. 6617
PID:20.500.12556/RUL-136425 This link opens in a new window
ISSN on article:1420-3049
DOI:10.3390/molecules26216617 This link opens in a new window
COBISS.SI-ID:83744003 This link opens in a new window
Publication date in RUL:03.05.2022
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Shortened title:Molecules
COBISS.SI-ID:18462981 This link opens in a 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:01.11.2021

Secondary language

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


Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:P4-0176
Name:Molekularna biotehnologija: od dinamike bioloških sistemov do aplikacij

Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:P4-0407
Name:Okoljska in aplikativna virologija: virusi, prijatelji in sovražniki

Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:P4-0165
Name:Biotehnologija in sistemska biologija rastlin

Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:P1-0208
Name:Farmacevtska kemija: načrtovanje, sinteza in vrednotenje učinkovin

Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:V4-2038
Name:DNK cepiva in peptidnih inhibitorji proti SARS-CoV-2

Funder:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Project number:I0-0022
Name:Mreža raziskovalnih infrastrukturnih centrov Univerze v Ljubljani (MRIC UL)

Funder:Drugi - Drug financer ali več financerjev
Funding programme:Ministry of Education, Science, and Sport (MIZŠ)
Project number:OP20.05187

Funder:EC - European Commission
Funding programme:European Regional Development Fund

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