VeroE6/TMPRSS2 cells (1.8 × 105) were seeded in a 24-well plate with culture medium (DMEM high glucose medium supplemented with 10% FCS, 1% L-Glutamine, 1% Penicillin/Streptomycin; all reagents were obtained from Sigma Aldrich, Missouri, USA) and incubated overnight at 37°C and 5% CO2. On the following day, whole serum or plasma samples were serial-diluted from 1:1 to 1:6,250 and incubated with SARS-CoV-2 strains (1.5 × 104 PFU/ml) for 1 h at 37°C. After incubation serum/plasma-virus mix was ultracentrifuged and resuspended in DMEM high glucose medium supplemented with 5% FCS, 1% L-Glutamine, 1% Penicillin/Streptomycin. VeroE6/TMPRSS2 cells were then inoculated with antibody-opsonized SARS-CoV-2 for 30 min on a shaker at RT and 30 min in an incubator at 37°C and 5% CO2. After incubation, inoculation medium was replaced with culture medium containing 1.5% Low-melt Agarose (Biozym, Oldendorf, Germany). Cells were incubated for 3 days at 37°C and 5% CO2 before plaque visualization and counting using 0.1% Neutral Red solution for 3 h (Sigma Aldrich, Missouri, USA).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.