20.500.12556/RUL-136425 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, 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. oral cavity swab passive drool pooling saliva COVID-19 SARS-CoV-2 LAMP RT-qPCR COVID-19 serological testing real-time polymerase chain reaction bris ustne sluznice združeni vzorci SARS-CoV-2 serološko testiranje za COVID-19 verižna reakcija s polimerazo v realnem času slina covid-19 true false true Angleški jezik Slovenski jezik Članek v reviji 2022-05-03 13:23:46 2022-05-03 13:23:51 2022-09-15 04:13:52 0000-00-00 00:00:00 2021 0 0 19 str. iss. 21, art. 6617 Vol. 26 Nov. 2021 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616.9 1420-3049 10.3390/molecules26216617 83744003 molecules-26-06617-v3.pdf molecules-26-06617-v3.pdf 1 18B9A7363F04D8631189C6EAF027EE7B cb7e08a4791135c5648a056589bd0289c622d8dd10d862d5be2b55eeef4a909a bd1862a7-cad3-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=156070 https://www.mdpi.com/1420-3049/26/21/6617 0 82f96a6c-cad3-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=156069 Fakulteta za farmacijo Biotehniška fakulteta 0 0 0