Detection of SARS-CoV-2 by RNAscope® in situ hybridization and immunohistochemistry techniques

Mariano Carossino; Hon S Ip; Juergen A Richt; Kendra Shultz; Kimberly Harper; Alan T Loynachan; Fabio Del Piero; Udeni B R Balasuriya

doi:10.1007/s00705-020-04737-w

Detection of SARS-CoV-2 by RNAscope^® in situ hybridization and immunohistochemistry techniques

Arch Virol. 2020 Oct;165(10):2373-2377. doi: 10.1007/s00705-020-04737-w. Epub 2020 Aug 5.

Authors

Mariano Carossino¹, Hon S Ip², Juergen A Richt³, Kendra Shultz¹, Kimberly Harper¹, Alan T Loynachan⁴, Fabio Del Piero¹, Udeni B R Balasuriya⁵

Affiliations

¹ Louisiana Animal Disease Diagnostic Laboratory, Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, River Rd, Room 1043, Baton Rouge, LA, 70803, USA.
² United States Department of the Interior, U. S. Geological Survey, National Wildlife Health Center, 6006, Schroeder Road, Madison, WI, 53711, USA.
³ Department of Diagnostic Medicine/Pathobiology, Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD), Kansas State University, K-224B Mosier Hall, Manhattan, KS, 66506, USA.
⁴ University of Kentucky Veterinary Diagnostic Laboratory, Department of Veterinary Science, University of Kentucky, Lexington, KY, 40511, USA.
⁵ Louisiana Animal Disease Diagnostic Laboratory, Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, River Rd, Room 1043, Baton Rouge, LA, 70803, USA. balasuriya1@lsu.edu.

Abstract

In situ hybridization (ISH) and immunohistochemistry (IHC) are essential tools to characterize SARS-CoV-2 infection and tropism in naturally and experimentally infected animals and also for diagnostic purposes. Here, we describe three RNAscope^®-based ISH assays targeting the ORF1ab, spike, and nucleocapsid genes and IHC assays targeting the spike and nucleocapsid proteins of SARS-CoV-2.

MeSH terms

Animals
Antibodies, Monoclonal
Antibodies, Viral
Antisense Elements (Genetics) / genetics
Betacoronavirus / genetics*
Betacoronavirus / isolation & purification*
COVID-19
COVID-19 Testing
Chlorocebus aethiops
Clinical Laboratory Techniques / methods*
Coronavirus Infections / diagnosis*
Coronavirus Infections / virology
Coronavirus Nucleocapsid Proteins
Genes, Viral
Humans
Immunohistochemistry / methods
In Situ Hybridization / methods
Nucleocapsid Proteins / genetics
Nucleocapsid Proteins / metabolism
Pandemics
Phosphoproteins
Pneumonia, Viral / diagnosis*
Pneumonia, Viral / virology
Polyproteins
RNA, Viral / genetics*
RNA, Viral / metabolism
SARS-CoV-2
Spike Glycoprotein, Coronavirus / genetics
Spike Glycoprotein, Coronavirus / metabolism
Vero Cells
Viral Proteins / genetics
Viral Proteins / metabolism

Substances

Antibodies, Monoclonal
Antibodies, Viral
Antisense Elements (Genetics)
Coronavirus Nucleocapsid Proteins
Nucleocapsid Proteins
ORF1ab polyprotein, SARS-CoV-2
Phosphoproteins
Polyproteins
RNA, Viral
Spike Glycoprotein, Coronavirus
Viral Proteins
nucleocapsid phosphoprotein, SARS-CoV-2
spike protein, SARS-CoV-2

Grants and funding

PG 002165/LSU Health Shreveport (US)