Exp Neurobiol.  2021 Feb;30(1):13-31. 10.5607/en20063.

Ultimate COVID-19 Detection Protocol Based on Saliva Sampling and qRT-PCR with Risk Probability Assessment

  • 1Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 34141, Korea
  • 2Center for Cognition and Sociality, Cognitive Glioscience Group, Institute for Basic Science, Daejeon 34126, Korea
  • 3Department of Biological Sciences, Middle East Technical University, Ankara 06800, Turkey
  • 4KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 02841, Korea
  • 5Department of Medical Genetics, Gazi University Faculty of Medicine, Ankara 06560, Turkey
  • 6Department of Chemistry, Middle East Technical University, Ankara 06800, Turkey
  • 7Department of Otorhinolaryngology, Seoul National University Bundang Hospital, Seongnam 13620, Korea


In the era of COVID-19 outbreak, various efforts are undertaken to develop a quick, easy, inexpensive, and accurate way for diagnosis. Although many commercial diagnostic kits are available, detailed scientific evaluation is lacking, making the public vulnerable to fear of false-positive results. Moreover, current tissue sampling method from respiratory tract requires personal contact of medical staff with a potential asymptomatic SARSCOV-2 carrier and calls for safe and less invasive sampling method. Here, we have developed a convenient detection protocol for SARS-COV-2 based on a non-invasive saliva self-sampling method by extending our previous studies on development of a laboratory-safe and low-cost detection protocol based on qRT-PCR. We tested and compared various self-sampling methods of self-pharyngeal swab and self-saliva sampling from non-carrier volunteers. We found that the self-saliva sampling procedure gave expected negative results from all of the non-carrier volunteers within 2 hours, indicating cost-effectiveness, speed and reliability of the saliva-based method. For an automated assessment of the sampling quality and degree of positivity for COVID-19, we developed scalable formulae based on a logistic classification model using both cycle threshold and melting temperature from the qRT-PCR results. Our newly developed protocol will allow easy sampling and spatial-separation between patient and experimenter for guaranteed safety. Furthermore, our newly established risk assessment formula can be applied to a large-scale diagnosis in health institutions and agencies around the world.


Coronavirus; COVID-19; SARS-CoV-2; Saliva sampling; COVID-19 risk assessment
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