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…
Divulgations
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 |
Tags
Citer Cet Article
Assessing the Effectiveness of an Antiviral Test Compound through a Viral Inactivation Assay. J. Vis. Exp. (Pending Publication), e21998, doi: (2024).