RT Journal Article
SR Electronic
T1 Molecular Detection of SARS-CoV-2 in Formalin Fixed Paraffin Embedded Specimens
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.21.042911
DO 10.1101/2020.04.21.042911
A1 Jun Liu
A1 April M. Babka
A1 Brian J. Kearney
A1 Sheli R. Radoshitzky
A1 Jens H. Kuhn
A1 Xiankun Zeng
YR 2020
UL http://biorxiv.org/content/early/2020/04/21/2020.04.21.042911.abstract
AB 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 one million cases and tens of thousands of deaths. 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.Competing Interest StatementThe authors have declared no competing interest.