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Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses
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...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344879/ https://www.ncbi.nlm.nih.gov/pubmed/35930888 http://dx.doi.org/10.1016/j.ebiom.2022.104193 |
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author | Krasemann, Susanne Dittmayer, Carsten von Stillfried, Saskia Meinhardt, Jenny Heinrich, Fabian Hartmann, Kristin Pfefferle, Susanne Thies, Edda von Manitius, Regina Aschman, Tom Alex David Radke, Josefine Osterloh, Anja Schmid, Simone Buhl, Eva Miriam Ihlow, Jana Dubois, Frank Arnhold, Viktor Elezkurtaj, Sefer Horst, David Hocke, Andreas Timm, Sara Bachmann, Sebastian Corman, Victor Goebel, Hans-Hilmar Matschke, Jakob Stanelle-Bertram, Stephanie Gabriel, Gülsah Seilhean, Danielle Adle-Biassette, Homa Ondruschka, Benjamin Ochs, Matthias Stenzel, Werner Heppner, Frank L. Boor, Peter Radbruch, Helena Laue, Michael Glatzel, Markus |
author_facet | Krasemann, Susanne Dittmayer, Carsten von Stillfried, Saskia Meinhardt, Jenny Heinrich, Fabian Hartmann, Kristin Pfefferle, Susanne Thies, Edda von Manitius, Regina Aschman, Tom Alex David Radke, Josefine Osterloh, Anja Schmid, Simone Buhl, Eva Miriam Ihlow, Jana Dubois, Frank Arnhold, Viktor Elezkurtaj, Sefer Horst, David Hocke, Andreas Timm, Sara Bachmann, Sebastian Corman, Victor Goebel, Hans-Hilmar Matschke, Jakob Stanelle-Bertram, Stephanie Gabriel, Gülsah Seilhean, Danielle Adle-Biassette, Homa Ondruschka, Benjamin Ochs, Matthias Stenzel, Werner Heppner, Frank L. Boor, Peter Radbruch, Helena Laue, Michael Glatzel, Markus |
author_sort | Krasemann, Susanne |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9344879 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-93448792022-08-02 Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses Krasemann, Susanne Dittmayer, Carsten von Stillfried, Saskia Meinhardt, Jenny Heinrich, Fabian Hartmann, Kristin Pfefferle, Susanne Thies, Edda von Manitius, Regina Aschman, Tom Alex David Radke, Josefine Osterloh, Anja Schmid, Simone Buhl, Eva Miriam Ihlow, Jana Dubois, Frank Arnhold, Viktor Elezkurtaj, Sefer Horst, David Hocke, Andreas Timm, Sara Bachmann, Sebastian Corman, Victor Goebel, Hans-Hilmar Matschke, Jakob Stanelle-Bertram, Stephanie Gabriel, Gülsah Seilhean, Danielle Adle-Biassette, Homa Ondruschka, Benjamin Ochs, Matthias Stenzel, Werner Heppner, Frank L. Boor, Peter Radbruch, Helena Laue, Michael Glatzel, Markus eBioMedicine Articles 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. Elsevier 2022-08-02 /pmc/articles/PMC9344879/ /pubmed/35930888 http://dx.doi.org/10.1016/j.ebiom.2022.104193 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Articles Krasemann, Susanne Dittmayer, Carsten von Stillfried, Saskia Meinhardt, Jenny Heinrich, Fabian Hartmann, Kristin Pfefferle, Susanne Thies, Edda von Manitius, Regina Aschman, Tom Alex David Radke, Josefine Osterloh, Anja Schmid, Simone Buhl, Eva Miriam Ihlow, Jana Dubois, Frank Arnhold, Viktor Elezkurtaj, Sefer Horst, David Hocke, Andreas Timm, Sara Bachmann, Sebastian Corman, Victor Goebel, Hans-Hilmar Matschke, Jakob Stanelle-Bertram, Stephanie Gabriel, Gülsah Seilhean, Danielle Adle-Biassette, Homa Ondruschka, Benjamin Ochs, Matthias Stenzel, Werner Heppner, Frank L. Boor, Peter Radbruch, Helena Laue, Michael Glatzel, Markus Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses |
title | Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses |
title_full | Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses |
title_fullStr | Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses |
title_full_unstemmed | Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses |
title_short | Assessing and improving the validity of COVID-19 autopsy studies - A multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses |
title_sort | assessing and improving the validity of covid-19 autopsy studies - a multicentre approach to establish essential standards for immunohistochemical and ultrastructural analyses |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344879/ https://www.ncbi.nlm.nih.gov/pubmed/35930888 http://dx.doi.org/10.1016/j.ebiom.2022.104193 |
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