immunolabelling as indisputable proof of SARS-CoV-2 virions in infected tissue
Andreja Erman (Author), Karmen Wechtersbach (Author), Dane Velkavrh (Author), Jerica Pleško (Author), Maja Frelih (Author), Nika Kojc (Author)

Abstract

Background: There is increasing evidence that identification of SARS-CoV-2 virions by transmission electron microscopy could be misleading due to the similar morphology of virions and ubiquitous cell structures. This study thus aimed to establish methods for indisputable proof of the presence of SARS-CoV-2 virions in the observed tissue. Methods: We developed a variant of the correlative microscopy approach for SARS-CoV-2 protein identification using immunohistochemical labelling of SARS-CoV-2 proteins on light and electron microscopy levels. We also performed immunogold labelling of SARS-CoV-2 virions. Results: Immunohistochemistry (IHC) of SARS-CoV-2 nucleocapsid proteins and subsequent correlative microscopy undoubtedly proved the presence of SARS-CoV-2 virions in the analysed human nasopharyngeal tissue. The presence of SARS-CoV-2 virions was also confirmed by immunogold labelling for the first time. Conclusions: Immunoelectron microscopy is the most reliable method for distinguishing intracellular viral particles from normal cell structures of similar morphology and size as virions. Furthermore, we developed a variant of correlative microscopy that allows pathologists to check the results of IHC performed first on routinely used paraffin-embedded samples, followed by semithin, and finally by ultrathin sections. Both methodological approaches indisputably proved the presence of SARS-CoV-2 virions in cells.

Keywords

SARS-CoV-2;immunolabelling;microscopy;

Data

Language: English
Year of publishing:
Typology: 1.01 - Original Scientific Article
Organization: UL MF - Faculty of Medicine
UDC: 616.9
COBISS: 76924931 Link will open in a new window
ISSN: 1999-4915
Views: 102
Downloads: 27
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Other data

Secondary language: Slovenian
Secondary keywords: SARS-CoV-2;imuno označevanje;mikroskopija;
Type (COBISS): Article
Pages: str. 1-9
Volume: ǂVol. ǂ13
Issue: ǂiss. ǂ9
Chronology: 2021
DOI: 10.3390/v13091816
ID: 15078744