Infectious Disease Research
Virology
Herpes Virus Research and Development
Mechanism of Action Testing
- Variation in multiplicity of infection: Currently available for HSV-1, HSV-2, HCMV and VZV. Compounds are evaluated for activity when challenged with different amounts of virus ranging from very low to very high multiplicity of infection.
- Time of addition and time of removal: Compounds are added or removed from cultures at various times pre- and post-infection. By comparison with other known herpesvirus inhibitors, this allows one to determine the relative point in the virus life cycle that is being inhibited (immediate early, early, late functions, DNA polymerization, etc.). This is a relatively standard technique typically used early during the process of determining mechanism of action as it allows one to narrow in on a smaller target window of activity for further experimentation. It also allows for an easy way to determine if a compound is acting by a unique or novel mechanism compared to other known inhibitors. Furthermore, time of removal studies allow one to determine the reversibility of a compounds activity.
- Analysis of viral DNA: The effect of compounds on the production of viral DNA can be evaluated using various hybridization techniques, PCR or TaqMan PCR.
- Analysis of viral proteins: The effect of compounds on the production of immediate early, early and late viral proteins can be evaluated using Western blots and/or Flow cytometry.
- Selection and characterization of drug-resistant virus isolates: Resistant virus isolates are selected in tissue culture by serial passage of the virus in the presence of gradually increasing concentrations of the compound. Resistance evaluations can be performed in any of the available cell lines with a variety of virus isolates. In addition, resistance selection can be evaluated using combinations of anti-HSV agents to evaluate the relative ability of the virus to become resistant to multiple agents that might be used in the clinic. Furthermore, these studies can be performed as part of a directed research project to determine the location of the drug resistance mutations that develop in the viral genome and transfer of the mutations into susceptible virus strains to demonstrate they are responsible for drug resistance (Marker Transfer Experiments).
- Directed research: Unique, directed research projects can be designed and implemented in order to further study the effects of a specific compound or class of compounds on herpesvirus replication. Alternatively, novel assays can be developed based upon identified herpesvirus drug targets and used to screen compound libraries for specific inhibitors of that target.
Primary Drug Screening Assays
Secondary Testing
Mechanism of Action Testing
Herpesvirus Web Resources
