| Autor: |
S Krasemann, C Dittmayer, S v. Stillfried, J Meinhardt, F Heinrich, K Hartmann, S Pfefferle, E Thies, R v. Manitius, T Aschman, J Radke, A Osterloh, S Schmid, EM Buhl, J Ihlow, S Elezkurtaj, D Horst, AC Hocke, S Timm, S Bachmann, V Corman, HH Goebel, J Matschke, S Stanelle-Bertram, G Gabriel, D Seilhean, H Adle-Biassette, B Ondruschka, M Ochs, W Stenzel, FL Heppner, P Boor, H Radbruch, M Laue, M Glatzel |
| Rok vydání: |
2022 |
| Předmět: |
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| DOI: |
10.1101/2022.01.13.22269205 |
| Popis: |
BackgroundAutopsy studies have provided valuable insights into the pathophysiology of COVID-19. Controversies remain whether the clinical presentation is due to direct organ damage by SARS-CoV-2 or secondary effects, e.g. by an 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.MethodsWe 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 spike protein and nucleocapsid, dsRNA, and non-structural protein Nsp3 in cultured cell lines and COVID-19 autopsy tissues. In a multicenter study, we evaluated specificity, reproducibility, and inter-observer variability of SARS-CoV-2 nucleocapsid staining. 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.FindingsPublications show high variability in the detection and interpretation of SARS-CoV-2 abundance in autopsy tissues by IHC or EM. In our study, 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 (r=-0.83, p-value InterpretationSince 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.Key messagesDetection of SARS-CoV-2 proteins by IHC in autopsy tissues is less sensitive in comparison to SARS-CoV-2 RNA detection by RT-qPCR.For determination of SARS-CoV-2 protein positive cells by IHC in autopsy tissues, detection of spike protein is less sensitive than nucleocapsid protein.Correct identification of SARS-CoV-2 particles in human samples by EM is limited to the respiratory system.Interpretation of IHC and EM should follow substantiated consensus criteria to enhance accuracy.Existing datasets describing SARS-CoV-2 presence in human autopsy tissues need to be critically re-evaluated. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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