Virus Plaque Assay: A Technique to Measure Infectious Herpes Simplex Virus via Quantification of Plaques Formed in Monolayer of Virus-Infected Cells
Published: April 30, 2023
Abstract
Source: Sadowski, L. A. et al. Plaquing of Herpes Simplex Viruses. J. Vis. Exp. (2021)
This video describes the virus plaque assay of the infectious herpes simplex virus or HSV. The diluted viral particles are inoculated to the monolayer of epithelial cells. Upon incubation, the virus forms plaques and is counted visually to determine the infectious virus titer.
Protocol
1. Seeding of the Vero cells The day before initiating the plaque assay, trypsinize Vero cells and resuspend them in regular Dulbecco's Modified Eagles Medium (DMEM) and supplement as per standard cell culture methodology. Resuspend the trypsinized cells in 10 mL of DMEM per T-75 flask of the confluent Vero cells (~107 cells). NOTE: DMEM is used for plaque assays instead of alpha-MEM because it slightly slows the cell growth rate and favors better plaque assay results.</…
Divulgazioni
The authors have nothing to disclose.
Materials
| 12-well plates | Corning | 3512 | |
| 6-well plates | Corning | 3516 | |
| Alpha-MEM | Lonza | 12169F | |
| Antibiotic/antimycotic | Gibco | 15240096 | |
| Crystal violet | Alfa Aesar | B2193214 | |
| DMEM | Gibco | 11965092 | |
| Dulbecco's PBS (no Mg++ or Ca++) | Gibco | 14190144 | |
| Fetal calf serum | Millipore-Sigma | TMS-013-B | |
| L-alanyl-L-glutamine (Glutamax) | Gibco | GS07F161BA | |
| Hemacytometer | Thermo Fisher | 02-671-54 | |
| Methylcellulose | Millipore-Sigma | 27-441-0 | |
| Quaternary agent (Lysol I.C.) | Thermo Fisher | NC9645698 | |
| Trypan Blue | Corning | 25900CI | |
| Trypsin | Cytiva | SH30042.01 | |
| Vero cells | ATCC | CCL-81 |
Tags
Citazione di questo articolo
Virus Plaque Assay: A Technique to Measure Infectious Herpes Simplex Virus via Quantification of Plaques Formed in Monolayer of Virus-Infected Cells. J. Vis. Exp. (Pending Publication), e21232, doi: (2023).