Assessing the Effectiveness of an Antiviral Test Compound through a Viral Inactivation Assay

Published: February 29, 2024

Abstract

Source: Tai, C. et al., Early Viral Entry Assays for the Identification and Evaluation of Antiviral Compounds. J. Vis. Exp. (2015)

This video demonstrates the viral inactivation assay, showcasing the evaluation of the impact of antiviral agents on viral particles.

Protocol

1. Viral Inactivation Assay Note: Examples of incubation period and viral dose for various viruses are listed in Figure 1A. Higher concentrations of the virus can also be tested by increasing the MOI/PFU. Seed Huh-7.5 cells in a 96-well plate (1 × 104 cells per well) and incubate at 37 °C in a 5% CO2 incubator O/N to obtain a monolayer. Incubate the test compounds or controls (final concentrations are: CHLA…

Representative Results

Figure 1. Inactivation of viral infections by the test compounds CHLA and PUG. Different viruses were treated with the test compounds for a long period (incubated for 1.5 – 3 hr before titration; light gray bars) or short period (immediately diluted; dark gray bars) at 37 °C before a dilution to sub-therapeutic concentration and subsequent analysis of infection on the respective host c…

Disclosures

The authors have nothing to disclose.

Materials

DMEM GIBCO 11995-040
FBS GIBCO 26140-079
Penicillin-Streptomycin GIBCO 15070-063
Amphotericin B GIBCO 15290-018
DMSO Sigma D5879
In vitro toxicology assay kit, XTTbased Sigma TOX2
PBS pH 7.4 GIBCO 10010023
Microplate reader Thermo Scientific 89087-320
Microcentrifuge Thermo Scientific 75002420
BioLux Gaussia luciferase assay kit New England Biolabs E3300L
Luminometer Promega GloMax-20/20
Sodium citrate, dihydrate Sigma 71402
Potassium chloride Sigma P5405

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Cite This Article
Assessing the Effectiveness of an Antiviral Test Compound through a Viral Inactivation Assay. J. Vis. Exp. (Pending Publication), e21998, doi: (2024).

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