Just seeing is not enough for believing : immunolabelling as indisputable proof of SARS-CoV-2 virions in infected tissue

Erman, Andreja (avtor), Wechtersbach, Karmen (avtor), Velkavrh, Dane (avtor), Pleško, Jerica (avtor), Frelih, Maja (avtor), Kojc, Nika (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: SARS-CoV-2, immunolabelling, microscopy, light microscopy, electron microscopy, correlative microscopy
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: MF - Medicinska fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2021
• Št. strani: 9 str.
• Številčenje: Vol. 13, iss. 9, art. 1816
• PID: 20.500.12556/RUL-136194
• UDK: 616.9
• ISSN pri članku: 1999-4915
• DOI: 10.3390/v13091816
• COBISS.SI-ID: 76924931
• Datum objave v RUL: 19.04.2022

Gradivo je del revije

• Naslov: Viruses
• Skrajšan naslov: Viruses
• Založnik: MDPI
• ISSN: 1999-4915
• COBISS.SI-ID: 517597977

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.
• Začetek licenciranja: 13.09.2021

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: SARS-CoV-2, imunooznačevanje, mikroskopija