SARS-CoV-2 Variant Growth Kinetics

SARS-CoV-2 USA-WA-1/2020 (NR-52281) (WA-1), B.1.1.7/Alpha (NR-54000), and B.1.351/Beta (NR-54008) strains were obtained from BEI Resources, and SARS-CoV-2 Delta variant B.1.617.2 hCoV-19/USA/WV-WVU-WV118685/2021 (GISAID Accession ID: EPI_ISL_1742834) was obtained from a patient sample at WVU. These strains were propagated in Vero E6 cells (ATCC-CRL-1586) as described (26 (link), 55 (link)). Vero E6 cells for viral titrations (6-well plate, 10⁶ cells/well) were infected with 10-fold serial dilutions of SARS-CoV-2. At 72 h post infection, cells were fixed overnight with 10% formalin (Sigma HT501128-4L), permeabilized and immunostained with 1 μg/ml of a SARS-CoV cross-reactive N protein antibody 1C7C7, kindly provided by Dr. Thomas Moran at the Icahn School of Medicine at Mount Sinai. For viral growth kinetics, Vero E6 cells (6-well plate, 10⁶ cells/well, triplicates) were infected (multiplicity of infection, MOI 0.01) with SARS-CoV-2 WA-1, Alpha or Beta. At the indicated times after viral infection (12, 24, 48 and 72 h), tissue culture samples were collected and titrated by plaque assay as described (26 (link)).

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Wong T.Y., Horspool A.M., Russ B.P., Ye C., Lee K.S., Winters M.T., Bevere J.R., Miller O.A., Rader N.A., Cooper M., Kieffer T., Sourimant J., Greninger A.L., Plemper R.K., Denvir J., Cyphert H.A., Barbier M., Torrelles J.B., Martinez I., Martinez-Sobrido L, & Damron F.H. (2022). Evaluating Antibody Mediated Protection against Alpha, Beta, and Delta SARS-CoV-2 Variants of Concern in K18-hACE2 Transgenic Mice. Journal of Virology, 96(6), e02184-21.

Publication 2022

SARS-CoV-2 USA-WA-1/2020 ( Vero E6 cells HT501128-4L

Antibody Assay B 1 1 7 B 1 351 Cells Cross reactive Dilutions Formalin Infection Kinetics Patient Plaque Sars cov 1 Sars cov 2 Sars cov 2 delta variant Sars cov n protein Strains Tissue Titrations Vero cells Viral infection

Corresponding Organization : West Virginia University

Other organizations : Texas Biomedical Research Institute, Georgia State University, University of Washington, Marshall University

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Variable analysis

independent variables

SARS-CoV-2 strains: USA-WA-1/2020 (NR-52281) (WA-1), B.1.1.7/Alpha (NR-54000), B.1.351/Beta (NR-54008), and B.1.617.2/Delta variant B.1.617.2 hCoV-19/USA/WV-WVU-WV118685/2021 (GISAID Accession ID: EPI_ISL_1742834)

dependent variables

Viral titers measured by plaque assay at 12, 24, 48, and 72 hours post-infection

control variables

Vero E6 cells (ATCC-CRL-1586) used for viral propagation and titrations
Cell culture conditions (6-well plate, 10^6 cells/well)
Viral infection conditions (multiplicity of infection, MOI 0.01)

controls

Positive control: SARS-CoV cross-reactive N protein antibody 1C7C7 used for immunostaining
Negative control: Not explicitly mentioned

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