Direct detection of SARS-CoV-2 antisense and sense genomic RNA in human saliva by semi-autonomous fluorescence in situ hybridization: A proxy for contagiousness?

Gijsbert J Jansen, Marit Wiersma, Willem J B van Wamel, Inge D Wijnberg

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Saliva is a matrix which may act as a vector for pathogen transmission and may serve as a possible proxy for SARS-CoV-2 contagiousness. Therefore, the possibility of detection of intracellular SARS-CoV-2 in saliva by means of fluorescence in situ hybridization is tested, utilizing probes targeting the antisense or sense genomic RNA of SARS-CoV-2. This method was applied in a pilot study with saliva samples collected from healthy persons and those presenting with mild or moderate COVID-19 symptoms. In all participants, saliva appeared a suitable matrix for the detection of SARS-CoV-2. Among the healthy, mild COVID-19-symptomatic and moderate COVID-19-symptomatic persons, 0%, 90% and 100% tested positive for SARS-CoV-2, respectively. Moreover, the procedure allows for simultaneous measurement of viral load ('presence', sense genomic SARS-CoV-2 RNA) and viral replication ('activity', antisense genomic SARS-CoV-2 RNA) and may yield qualitative results. In addition, the visualization of DNA in the cells in saliva provides an additional cytological context to the validity and interpretability of the test results. The method described in this pilot study may be a valuable diagnostic tool for detection of SARS-CoV-2, distinguishing between 'presence' (viral load) and 'activity' (viral replication) of the virus. Moreover, the method potentially gives more information about possible contagiousness.

Original languageEnglish
Article numbere0256378
Pages (from-to)1-11
Number of pages11
JournalPLoS One
Volume16
Issue number8
DOIs
Publication statusPublished - 17 Aug 2021

Bibliographical note

Publisher Copyright:
© 2021 Jansen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Keywords

  • COVID-19/diagnosis
  • Case-Control Studies
  • Genomics
  • Humans
  • In Situ Hybridization, Fluorescence/methods
  • RNA, Antisense/genetics
  • RNA, Viral/analysis
  • SARS-CoV-2/genetics
  • Saliva/virology
  • Severity of Illness Index
  • Viral Load
  • Virus Replication

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