학술논문
Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses
Document Type
article
Author
Susanne Krasemann; Carsten Dittmayer; Saskia von Stillfried; Jenny Meinhardt; Fabian Heinrich; Kristin Hartmann; Susanne Pfefferle; Edda Thies; Regina von Manitius; Tom Alex David Aschman; Josefine Radke; Anja Osterloh; Simone Schmid; Eva Miriam Buhl; Jana Ihlow; Frank Dubois; Viktor Arnhold; Sefer Elezkurtaj; David Horst; Andreas Hocke; Sara Timm; Sebastian Bachmann; Victor Corman; Hans-Hilmar Goebel; Jakob Matschke; Stephanie Stanelle-Bertram; Gülsah Gabriel; Danielle Seilhean; Homa Adle-Biassette; Benjamin Ondruschka; Matthias Ochs; Werner Stenzel; Frank L. Heppner; Peter Boor; Helena Radbruch; Michael Laue; Markus Glatzel
Source
EBioMedicine, Vol 83, Iss , Pp 104193- (2022)
Subject
Language
English
ISSN
2352-3964
Abstract
Summary: Background: Autopsy studies have provided valuable insights into the pathophysiology of COVID-19. Controversies remain about whether the clinical presentation is due to direct organ damage by SARS-CoV-2 or secondary effects, such as overshooting immune response. SARS-CoV-2 detection in tissues by RT-qPCR and immunohistochemistry (IHC) or electron microscopy (EM) can help answer these questions, but a comprehensive evaluation of these applications is missing. Methods: We assessed publications using IHC and EM for SARS-CoV-2 detection in autopsy tissues. We systematically evaluated commercially available antibodies against the SARS-CoV-2 proteins in cultured cell lines and COVID-19 autopsy tissues. In a multicentre study, we evaluated specificity, reproducibility, and inter-observer variability of SARS-CoV-2 IHC. We correlated RT-qPCR viral tissue loads with semiquantitative IHC scoring. We used qualitative and quantitative EM analyses to refine criteria for ultrastructural identification of SARS-CoV-2. Findings: Publications show high variability in detection and interpretation of SARS-CoV-2 abundance in autopsy tissues by IHC or EM. We show that IHC using antibodies against SARS-CoV-2 nucleocapsid yields the highest sensitivity and specificity. We found a positive correlation between presence of viral proteins by IHC and RT-qPCR-determined SARS-CoV-2 viral RNA load (N= 35; r=-0.83, p-value