Antiviral susceptibility testing with a cell line which expresses beta-galactosidase after infection with herpes simplex virus
Open Access
- 1 June 1995
- journal article
- research article
- Published by American Society for Microbiology in Antimicrobial Agents and Chemotherapy
- Vol. 39 (6) , 1287-1291
- https://doi.org/10.1128/aac.39.6.1287
Abstract
Despite increasing concern about drug-resistant herpes simplex virus (HSV), antiviral susceptibility testing is not routinely performed by most clinical virology laboratories. This omission is in large part because the most widely accepted method, the plaque reduction assay (PRA), is cumbersome to perform and results are rarely available in time to influence treatment. We report here the development of a sensitivity test for HSV which utilizes a cell line (VeroICP6LacZ#7) that expresses beta-galactosidase activity after infection with HSV such that infected cells can be detected by histochemical staining. We designed an assay in which 10-fold dilutions of virus stocks with undetermined titers were inoculated onto VeroICP6LacZ#7 cells in a 24-well tissue culture dish. Forty-eight hours after infection, the cell monolayers were histochemically stained. Plaques appear blue against a clear background and are thus easily visualized at 48 h. As with the standard PRA, the 50% inhibitory concentration (IC50) was reported as the concentration of an antiviral drug that reduces the number of plaques by 50%. Evaluation of 10 well-characterized laboratory strains and 12 clinical HSV isolates showed that the IC50 determined by this method correlated in all instances with the IC50 determined by the PRA. This method is easy to use and eliminates the need to determine the titer of the virus, and results are available within 48 h of the detection of the virus. VeroICP6Lac#7 cells are a useful tool for performing HSV antiviral susceptibility testing and could be used in a number of different formats to facilitate the identification of drug-resistant isolates of HSV.Keywords
This publication has 22 references indexed in Scilit:
- A rapid and reliable assay for testing acyclovir sensitivity of clinical herpes simplex virus isolates independent of virus dose and reading timeAntiviral Research, 1994
- Foscarnet-Resistant Herpes Simplex Virus Infection in Patients with AIDSThe Journal of Infectious Diseases, 1994
- The herpes simplex virus type 1 ICP6 gene is regulated by a ‘leaky’ early promoterVirus Research, 1992
- Isolation of a cell line for rapid and sensitive histochemical assay for the detection of herpes simplex virusJournal of Virological Methods, 1992
- A Controlled Trial Comparing Foscarnet with Vidarabine for Acyclovir-Resistant Mucocutaneous Herpes Simplex in the Acquired Immunodeficiency SyndromeNew England Journal of Medicine, 1991
- Simplified test for detecting the resistance of herpes simplex virus to acyclovirJournal of Medical Virology, 1990
- Susceptibility to acyclovir of herpes simplex virus: Emergence of resistance in patients with lymphoid and myeloid neoplasiaJournal of Infection, 1989
- Acyclovir-Resistant Herpes Simplex Virus Infections in Patients with the Acquired Immunodeficiency SyndromeNew England Journal of Medicine, 1989
- A colorimetric assay for the measurement of the sensitivity of herpes simplex viruses to antiviral agentsAntiviral Research, 1983
- Trisodium Phosphonoformate, a New Antiviral CompoundScience, 1978