Go to The Journal of Clinical Investigation
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Immunology
    • Metabolism
    • Nephrology
    • Oncology
    • Pulmonology
    • All ...
  • Videos
  • Collections
    • Resource and Technical Advances
    • Clinical Medicine
    • Reviews
    • Editorials
    • Perspectives
    • Top read articles
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • In-Press Preview
  • Concise Communication
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
Molecular detection of SARS-CoV-2 in formalin-fixed, paraffin-embedded specimens
Jun Liu, … , Jens H. Kuhn, Xiankun Zeng
Jun Liu, … , Jens H. Kuhn, Xiankun Zeng
Published May 7, 2020
Citation Information: JCI Insight. 2020;5(12):e139042. https://doi.org/10.1172/jci.insight.139042.
View: Text | PDF
Resource and Technical Advance COVID-19 Virology

Molecular detection of SARS-CoV-2 in formalin-fixed, paraffin-embedded specimens

  • Text
  • PDF
Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China, in December 2019. The virus rapidly spread globally, resulting in a public health crisis including almost 5 million cases and 323,256 deaths as of May 21, 2020. Here, we describe the identification and evaluation of commercially available reagents and assays for the molecular detection of SARS-CoV-2 in infected FFPE cell pellets. We identified a suitable rabbit polyclonal anti–SARS-CoV spike protein antibody and a mouse monoclonal anti–SARS-CoV nucleocapsid protein (NP) antibody for cross-detection of the respective SARS-CoV-2 proteins by IHC and immunofluorescence assay (IFA). Next, we established RNAscope in situ hybridization (ISH) to detect SARS-CoV-2 RNA. Furthermore, we established a multiplex FISH (mFISH) to detect positive-sense SARS-CoV-2 RNA and negative-sense SARS-CoV-2 RNA (a replicative intermediate indicating viral replication). Finally, we developed a dual staining assay using IHC and ISH to detect SARS-CoV-2 antigen and RNA in the same FFPE section. It is hoped that these reagents and assays will accelerate COVID-19 pathogenesis studies in humans and in COVID-19 animal models.

Authors

Jun Liu, April M. Babka, Brian J. Kearney, Sheli R. Radoshitzky, Jens H. Kuhn, Xiankun Zeng

×

Figure 3

Detection of SARS-CoV-2 replication in FFPE cells using mFISH.

Options: View larger image (or click on image) Download as PowerPoint
Detection of SARS-CoV-2 replication in FFPE cells using mFISH.
(A and B)...
(A and B) Compared with uninfected control (A), SARS-CoV-2 negative-sense RNA (green), a replicative intermediate that indicates viral replication, can be detected in infected FFPE cell pellets in addition to positive-sense (red) RNA (B). Nuclei are stained blue (DAPI). Scale bars: 20 μm.

Copyright © 2022 American Society for Clinical Investigation
ISSN 2379-3708

Sign up for email alerts