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

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

doi:10.1172/jci.insight.139042

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

Jun Liu,¹ April M. Babka,¹ Brian J. Kearney,² Sheli R. Radoshitzky,³ Jens H. Kuhn,⁴ and Xiankun Zeng¹

Published May 7, 2020 - More info

View 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 more than 3.1 million cases and 224,000 deaths as of May 1, 2020. Here, we describe the identification and evaluation of commercially available reagents and assays for the molecular detection of SARS-CoV-2 in infected formalin fixed paraffin embedded (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 immunohistochemistry (IHC) and immunofluorescence assay (IFA). Next, we established RNAscope in situ hybridization (ISH) to detect SARS-CoV-2 RNA. Furthermore, we established a multiplex fluorescence ISH (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. These reagents and assays will accelerate COVID-19 pathogenesis studies in humans and in COVID-19 animal models.

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

Jun Liu,¹ April M. Babka,¹ Brian J. Kearney,² Sheli R. Radoshitzky,³ Jens H. Kuhn,⁴ and Xiankun Zeng¹

Article tools

Metrics

Go to

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

Jun Liu,1 April M. Babka,1 Brian J. Kearney,2 Sheli R. Radoshitzky,3 Jens H. Kuhn,4 and Xiankun Zeng1

Article tools

Metrics

Go to

Sign up for email alerts

Jun Liu,¹ April M. Babka,¹ Brian J. Kearney,² Sheli R. Radoshitzky,³ Jens H. Kuhn,⁴ and Xiankun Zeng¹