Este artigo é um Preprint
Preprints são relatos preliminares de pesquisa que não foram certificados pela revisão por pares. Eles não devem ser considerados para orientar a prática clínica ou comportamentos relacionados à saúde e não devem ser publicados na mídia como informação estabelecida.
Preprints publicados online permitem que os autores recebam feedback rápido, e toda a comunidade científica pode avaliar o trabalho independentemente e responder adequadamente. Estes comentários são publicados juntamente com os preprints para qualquer pessoa ler e servir como uma avaliação pós-publicação.
Intracellular flow cytometry complements RT-qPCR detection of circulating SARS-CoV-2 variants of concern
Preprint
em En
| PREPRINT-BIORXIV
| ID: ppbiorxiv-478775
ABSTRACT
Despite the efficacy of current vaccines against SARS-CoV-2, the spread of the virus is still not under control, as evidenced by the ongoing circulation of the highly contagious SARS-CoV-2 Omicron variant. Basic and antiviral research on SARS-CoV-2 relies 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 RT-qPCR detection and can be applied to all current SARS-CoV-2 variants of concern, including the highly mutated Omicron variant. Method summaryThis study describes the characterization of SARS-CoV-2 infected cells using intracellular flow cytometric viral nucleocapsid staining that complements RT-qPCR quantification of viral RNA. The technique makes it possible to distinguish between infected cells in the early (low N) or late phase (high N) of viral replication. It can also be applied to the different variants of concern of SARS-CoV-2, including the Omicron variant.
cc_no
Texto completo:
1
Coleções:
09-preprints
Base de dados:
PREPRINT-BIORXIV
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Preprint