Dataset Information

Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses.

ABSTRACT:

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.

SUBMITTER: Krasemann S

PROVIDER: S-EPMC9344879 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Publications

Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses.

Krasemann Susanne S Dittmayer Carsten C von Stillfried Saskia S Meinhardt Jenny J Heinrich Fabian F Hartmann Kristin K Pfefferle Susanne S Thies Edda E von Manitius Regina R Aschman Tom Alex David TAD Radke Josefine J Osterloh Anja A Schmid Simone S Buhl Eva Miriam EM Ihlow Jana J Dubois Frank F Arnhold Viktor V Elezkurtaj Sefer S Horst David D Hocke Andreas A Timm Sara S Bachmann Sebastian S Corman Victor V Goebel Hans-Hilmar HH Matschke Jakob J Stanelle-Bertram Stephanie S Gabriel Gülsah G Seilhean Danielle D Adle-Biassette Homa H Ondruschka Benjamin B Ochs Matthias M Stenzel Werner W Heppner Frank L FL Boor Peter P Radbruch Helena H Laue Michael M Glatzel Markus M

EBioMedicine 20220802

<h4>Background</h4>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.<h4>Methods</h4>We assessed pub ...[more]

PMID: 35930888

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Dataset Information

Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses.

Background

Methods

Findings

Interpretation

Funding

Publications

Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses.

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