Intracellular flow cytometry complements RT-qPCR detection of circulating SARS-CoV-2 variants of concern

BioTechniques

Joren Stroobants¹  Kurt Vermeire¹  Becky Provinciael²  Piet Maes²  Emiel Vanhulle³  Anita Camps³ 

[1] Chemotherapy, Herestraat 49, Leuven, 3000, Belgium;;Transplantation, Rega Institute, Laboratory of Virology &;1KU Leuven, Department of Microbiology, Immunology &

关键词: antiviral drug;    duplex RT-qPCR;    flow cytometry;    nucleocapsid;    omicron variant;    SARS-CoV-2;   

DOI  :  10.2144/btn-2022-0018

来源: DOAJ

【 摘 要 】

Basic and antiviral research on SARS-CoV-2 rely on cellular assays of virus replication in vitro. In addition, accurate detection of virus-infected cells and released virus particles is needed to study virus replication and to profile new candidate antiviral drugs. Here, by flow cytometry, we detect SARS-CoV-2 infection at single cell level and distinguish infected Vero E6 cells from uninfected bystander cells. Furthermore, based on the viral nucleocapsid expression, subpopulations of infected cells that are in an early or late phase of viral replication can be differentiated. Importantly, this flow cytometric technique complements our duplex RT-qPCR detection of viral E and N, and it can be applied to all current SARS-CoV-2 variants of concern, including the highly mutated Omicron variant.

【 授权许可】

Unknown