Procedures for Flow Cytometry-Based Sorting of Unfixed Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infected Cells and Other Infectious Agents

Kristen M Reifel; Brandon K Swan; Evan R Jellison; David Ambrozak; Jan Baijer; Richard Nguyen; Simon Monard; Geoffrey Lyon; Benjamin Fontes; Stephen P Perfetto

doi:10.1002/cyto.a.24040

Procedures for Flow Cytometry-Based Sorting of Unfixed Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infected Cells and Other Infectious Agents

Cytometry A. 2020 Jul;97(7):674-680. doi: 10.1002/cyto.a.24040. Epub 2020 Jun 3.

Authors

Affiliations

¹ National Biodefense Analysis and Countermeasures Center, Frederick, Maryland, USA.
² Department of Immunology, UConn Health, Farmington, Connecticut, USA.
³ Vaccine Research Center, NIAID, NIH, Bethesda, Maryland, USA.
⁴ CEA-DSV-IRCM, Fontenay-aux-Roses, France.
⁵ Walter and Eliza Hall Institute, Parkville, Victoria, Australia.
⁶ Yale University Flow Cytometry Facility, New Haven, Connecticut, USA.
⁷ Yale University Environmental Health and Safety Office, New Haven, Connecticut, USA.

Abstract

In response to the recent COVID-19 pandemic, many laboratories are involved in research supporting SARS-CoV-2 vaccine development and clinical trials. Flow cytometry laboratories will be responsible for a large part of this effort by sorting unfixed antigen-specific lymphocytes. Therefore, it is critical and timely that we have an understanding of risk assessment and established procedures of infectious cell sorting. Here we present procedures covering the biosafety aspects of sorting unfixed SARS-CoV-2-infected cells and other infectious agents of similar risk level. These procedures follow the ISAC Biosafety Committee guidelines and were recently approved by the National Institutes of Health Institutional Biosafety Committee for sorting SARS-CoV-2-infected cells. © 2020 International Society for Advancement of Cytometry.

Keywords: COVID-19; ISAC Biosafety Committee; SARS-CoV-2 cell sorting procedure; biosafety and cell sorting; coronavirus; infectious cell sorting.

MeSH terms

Betacoronavirus / isolation & purification*
COVID-19
Containment of Biohazards / methods*
Coronavirus Infections / diagnosis
Coronavirus Infections / prevention & control*
Flow Cytometry / methods*
Humans
Laboratories / standards
Medical Laboratory Personnel / standards
Pandemics / prevention & control*
Pneumonia, Viral / diagnosis
Pneumonia, Viral / prevention & control*
Risk Assessment
SARS-CoV-2
Specimen Handling / methods*