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

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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).

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