EBioMedicine (Sep 2022)
Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses
- 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
Affiliations
- Susanne Krasemann
- Institute of Neuropathology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
- Carsten Dittmayer
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Saskia von Stillfried
- Institute of Pathology and Electron Microscopy Facility, RWTH University of Aachen, Germany
- Jenny Meinhardt
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Fabian Heinrich
- Institute of Legal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
- Kristin Hartmann
- Institute of Neuropathology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
- Susanne Pfefferle
- Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
- Edda Thies
- Institute of Neuropathology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
- Regina von Manitius
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Tom Alex David Aschman
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Josefine Radke
- Department of Pathology, Universitätsmedizin Greifswald, Greifswald, Germany
- Anja Osterloh
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Simone Schmid
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Eva Miriam Buhl
- Institute of Pathology and Electron Microscopy Facility, RWTH University of Aachen, Germany
- Jana Ihlow
- Institute for Pathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany
- Frank Dubois
- Institute for Pathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany
- Viktor Arnhold
- Institute for Pathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany
- Sefer Elezkurtaj
- Institute for Pathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany
- David Horst
- Institute for Pathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany
- Andreas Hocke
- Department of Infectious Diseases and Respiratory Medicine, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany
- Sara Timm
- Core Facility Electron Microscopy, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Sebastian Bachmann
- Institute of Functional Anatomy, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Victor Corman
- Institute of Virology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin, Germany
- Hans-Hilmar Goebel
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Jakob Matschke
- Institute of Neuropathology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
- Stephanie Stanelle-Bertram
- Department for Viral Zoonoses-One Health, Leibniz Institute for Experimental Virology, Hamburg, Germany
- Gülsah Gabriel
- Department for Viral Zoonoses-One Health, Leibniz Institute for Experimental Virology, Hamburg, Germany; Institute for Virology, University for Veterinary Medicine Hannover, Germany
- Danielle Seilhean
- Raymond Escourolle Department of Neuropathology, Pitié-Salpêtrière Hospital, APHP, Sorbonne University, Paris, France
- Homa Adle-Biassette
- Department of Pathology, Université de Paris, AP-HP, Lariboisière Hospital, DMU DREAM, UMR 1141, INSERM, Paris, France
- Benjamin Ondruschka
- Institute of Legal Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
- Matthias Ochs
- Core Facility Electron Microscopy, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; Institute of Functional Anatomy, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany; German center for lung research (DZL), Berlin, Germany
- Werner Stenzel
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Frank L. Heppner
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Peter Boor
- Institute of Pathology and Electron Microscopy Facility, RWTH University of Aachen, Germany
- Helena Radbruch
- Department of Neuropathology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany
- Michael Laue
- National Consultant Laboratory for Electron Microscopy of Infectious Pathogens, Centre for Biological Threats and Special Pathogens 4 (ZBS 4), Robert Koch Institute, Berlin, Germany
- Markus Glatzel
- Institute of Neuropathology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; Corresponding author at: Institute of Neuropathology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
- Journal volume & issue
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Vol. 83
p. 104193
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 <0.0001). For EM, we refined criteria for virus identification and provide recommendations for optimized sampling and analysis. 135 of 144 publications misinterpret cellular structures as virus using EM or show only insufficient data. We provide publicly accessible digitized EM sections as a reference and for training purposes. Interpretation: Since detection of SARS-CoV-2 in human autopsy tissues by IHC and EM is difficult and frequently incorrect, we propose criteria for a re-evaluation of available data and guidance for further investigations of direct organ effects by SARS-CoV-2. Funding: German Federal Ministry of Health, German Federal Ministry of Education and Research, Berlin University Alliance, German Research Foundation, German Center for Infectious Research.
