Propagation and Quantification of SARS-CoV-2

Alexander S. Jureka, Christopher F. Basler

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

In late 2019, the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, China. Since its emergence, SARS-CoV-2 has been responsible for a world-wide pandemic resulting in over 80 million infections and over 1.8 million deaths. The severity of the pandemic has prompted widespread research efforts to more fully understand SARS-CoV-2 and the disease it causes, COVID-19. Research into this novel virus will be facilitated by the availability of clearly described and effective protocols that enable the propagation and quantification of infectious virus. Here, we describe protocols for the propagation of SARS-CoV-2 in Vero E6 cells as well as two human cells lines, the intestinal epithelial Caco-2 cell line and the respiratory epithelial Calu-3 cell line. Additionally, we provide protocols for the quantification of SARS-CoV-2 by plaque assays and immunofocus forming assays in Vero E6 cells utilizing liquid overlays. These protocols provide a foundation for laboratories acquiring the ability to study SARS-CoV-2 to address this ongoing pandemic.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages111-129
Number of pages19
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2452
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • COVID-19
  • Coronavirus
  • Focus forming assay
  • Plaque assay
  • SARS-CoV-2
  • Virus

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