A Viral Attachment Assay for Screening of Antiviral Compounds

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 a viral attachment assay for screening antiviral compounds. Using genetically modified hepatitis C virus and luciferase, it assesses inhibitory effects on viral attachment, as indicated by lower light intensity in co-treated wells compared to wells treated with the virus alone.

Protocol

NOTE: Ensure that all procedures involving cell culture and virus infection are conducted in certified biosafety hoods that are appropriate for the biosafety level of the samples being handled. For the purpose of describing the protocols, Gaussia luciferase reporter-tagged hepatitis C virus (HCV) is used as a model virus. In the context of the representative results, the compounds chebulagic acid (CHLA) and punicalagin (PUG) are used as candidate antivirals that target viral glycoprotein interaction…

Representative Results

Figure 1. Evaluation of antiviral activities of the test compounds CHLA and PUG against virus attachment and entry/fusion. (A) The experimental procedure, virus concentration (PFU/well or MOI), and the time of addition and treatment with the test compounds (i, ii, iii) are presented for each virus in the schematics and the associated tables. In virus attachment analysis (light gray bars), mo…

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, XTT-based Sigma TOX2
PBS pH 7.4 GIBCO 10010023
Microplate reader Bio-Tek Instrument, Inc. ELx800
Microcentrifuge Thermo Scientific 75002420
BioLux Gaussia luciferase assay kit New England Biolabs E3300L
Luminometer Promega GloMax-20/20

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Citer Cet Article
A Viral Attachment Assay for Screening of Antiviral Compounds. J. Vis. Exp. (Pending Publication), e21999, doi: (2024).

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