title: A rapid assay for evaluation of antiviral activity against coxsackie virus B3, influenza virus A, and herpes simplex virus type 1.
abstract: 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.
authors.affiliation: Institute for Virology, Medical Centre of the Friedrich Schiller University, Winzerlaer Str. 10, D-07745, Jena, Germany. i6scmi@pop3.uni-jena.de
authors.forename: M
authors.initials: M
authors.lastname: Schmidtke
authors.forename: U
authors.initials: U
authors.lastname: Schnittler
authors.forename: B
authors.initials: B
authors.lastname: Jahn
authors.forename: H
authors.initials: H
authors.lastname: Dahse
authors.forename: A
authors.initials: A
authors.lastname: Stelzner
mesh_terms: D000547:Amantadine; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002455:Cell Division; D002460:Cell Line; D002522:Chlorocebus aethiops; D003588:Cytopathogenic Effect, Viral; D004285:Dogs; D029822:Enterovirus B, Human / Q000187:drug effects* / Q000472:pathogenicity; D017245:Foscarnet; D019791:Guanidine; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000472:pathogenicity; D006801:Humans; D009980:Influenza A virus / Q000187:drug effects* / Q000472:pathogenicity; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors; D010948:Viral Plaque Assay
chemical_list: D000998:Antiviral Agents; D017245:Foscarnet; D000547:Amantadine; D019791:Guanidine