A rapid assay for evaluation of antiviral activity against coxsackie virus B3, influenza virus A, and herpes simplex virus type 1

• Published in: Journal of virological methods 2001-06, Vol.95 (1), p.133-143
• Main Authors: Schmidtke, M, Schnittler, U, Jahn, B, Dahse, H.-M, Stelzner, A
• Format: Article
• Language: English
• Subjects: Amantadine; Animal viral diseases; Animals; Antiviral Agents - pharmacology; Antiviral screening; Biological and medical sciences; Cell Division; Cell Line; Cercopithecus aethiops; Coxsackie virus B3; Coxsackievirus B3; Cytopathogenic Effect, Viral; Dogs; Enterovirus B, Human - drug effects; Enterovirus B, Human - pathogenicity; Foscarnet; Fundamental and applied biological sciences. Psychology; gentian violet; Guanidine; HeLa Cells; herpes simplex virus 1; Herpesvirus 1, Human - drug effects; Herpesvirus 1, Human - pathogenicity; HSV-1; human coxsackievirus B3; Humans; Infectious diseases; influenza A virus; Influenza A virus - drug effects; Influenza A virus - pathogenicity; Influenza virus A; Medical sciences; Microbiology; phosphonoformic acid; Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains; Reproducibility of Results; Staining and Labeling - methods; Time Factors; Viral diseases; Viral Plaque Assay; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(01)00305-6

In order to identify new potential antiviral drugs, small amounts of extracts or compounds have to be examined for cytotoxicity and antiviral activity in primary screening using a rapid, easy, inexpensive, and highly standardised test system. In this study, high-throughput cytopathic effect (CPE) inhibitory assays were established for coxsackie virus B3 on HeLa Ohio cells, influenza virus A on Madin–Darby canine kidney cells, and herpes simplex virus type 1 (HSV-1) on green monkey kidney cells that meet these requirements. The cytotoxic and the antiviral effects were quantified using a crystal violet uptake assay allowing automated handling of large numbers of candidate agents. To ensure comparable results with plaque reduction assays, the 50 and 90% plaque inhibitory concentrations of guanidine, amantadine, and phosphonoformic acid were used to standardise the anti-coxsackie virus B3, anti-influenza virus A, and anti-HSV-1 tests, respectively. The strong correlation between the antiviral activity determined by CPE-inhibitory assays and plaque reduction assay was further proved for other antivirals. In summary, low amounts of large numbers of compounds may be tested inexpensively and standardised within 24 h (coxsackie virus B3 and influenza virus A) or 48 h (herpes simplex virus type 1) post-infection using CPE inhibitory assays.