JoVE Encyclopedia of Experiments
Microbiology
0 views • 4:56 min • September 30th, 2026
Take a multi-well plate with heat-inactivated patient serum containing coronavirus-specific antibodies in the test well. Include healthy human serum as the control.
Add pseudoviruses displaying coronavirus spike proteins and carrying green fluorescent protein-encoding RNA to the serum samples, and incubate.
In the test well, antibodies bind to the spike proteins and neutralize the pseudoviruses, while in the control well, nonspecific antibodies do not bind to the spike proteins, leaving the pseudoviruses unbound.
Add the mixture to epithelial cell monolayers. Incubate for viral adsorption.
Add semi-solid overlay media to each well to restrict viral diffusion. Incubate.
In the control, unbound pseudoviruses bind to cell receptors, enter cells, release RNA, and express GFP, forming fluorescent infection foci.
In the test well, antibody-neutralized pseudoviruses fail to enter cells, preventing infection foci formation.
Under a fluorescent imager, count the fluorescent infection foci.
Absence of infection foci in the test well, compared with numerous foci in the control, confirms antibody-mediated neutralization of pseudoviruses by the patient serum.
To plate cells for a neutralization assay, u
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