Concentration of infectious SARS-CoV-2 by polyethylene glycol precipitation

J Virol Methods. 2020 Dec:286:113977. doi: 10.1016/j.jviromet.2020.113977. Epub 2020 Sep 23.

Abstract

The development of medical countermeasures against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) requires robust viral assays. Here we have adapted a protocol for polyethylene glycol (PEG)-mediated precipitation of SARS-CoV-2 stocks without the need for ultracentrifugation. Virus precipitation resulted in a ∼1.5 log10 increase in SARS-CoV-2 titres of virus prepared in VeroE6 cells and enabled the infection of several immortalized human cell lines (Caco-2 and Calu-3) at a high multiplicity of infection not practically achievable without virus concentration. This protocol underscores the utility of PEG-mediated precipitation for SARS-CoV-2 and provides a resource for a range of coronavirus research areas.

Keywords: COVID-19; PEG precipitation; SARS-CoV-2; Virus concentration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Betacoronavirus / isolation & purification*
  • COVID-19
  • COVID-19 Testing
  • Caco-2 Cells
  • Chlorocebus aethiops
  • Clinical Laboratory Techniques / methods*
  • Coronavirus Infections / diagnosis
  • Coronavirus Infections / virology*
  • Humans
  • Pandemics
  • Pneumonia, Viral / diagnosis
  • Pneumonia, Viral / virology*
  • Polyethylene Glycols / chemistry*
  • Reverse Transcriptase Polymerase Chain Reaction
  • SARS-CoV-2
  • Ultracentrifugation / methods
  • Vero Cells

Substances

  • Polyethylene Glycols