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Alberto van Olphen, D.V.M., Ph.D. Center for Biological Defense

Alberto van Olphen, D.V.M., Ph.D. Center for Biological Defense Global Health Infectious Disease Research College of Public Health University of South Florida . Taylor made screening protocols. In Vitro evaluation of compounds with antiviral activity against Dengue .

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Alberto van Olphen, D.V.M., Ph.D. Center for Biological Defense

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  1. Alberto van Olphen, D.V.M., Ph.D. Center for Biological Defense Global Health Infectious Disease Research College of Public Health University of South Florida

  2. Taylor made screening protocols

  3. In Vitro evaluation of compounds with antiviral activity against Dengue • Determination of cytotoxicity of compounds in selected cell line • MTT assay • Specific cytotox panels

  4. In Vitro evaluation of compounds with antiviral activity against Dengue • Cell-based assays • Determination of the EC50 antiviral compound activity

  5. Cell line matters • Vero - Cercopithecus aethiops • Aedesalbopictus • LLC-MK2 – MacacaMulatta • Your choice

  6. Serotype matters • Dengue Type 1Hawaii TC adapted NR 82 • Dengue Type 2 Virus, Strain New Guinea C NR84 • Dengue Type 3 , Strain H87 NR-80 • Dengue Virus Type 4 BC123/97 NR-3803 • Field isolates Dengue virus type 1, (Florida Keys isolate)

  7. In Vitro evaluation of compounds with antiviral activity against Dengue • Determination of plaque formation characteristics of specified viral strains and field isolates to identify the best method for virus titration

  8. In Vitro evaluation of compounds with antiviral activity against Dengue Determination of virus optimal harvest time using a time course experiment.

  9. TCID50 using immunostaining for non-cytopathic viruses

  10. Plaque assay – Focus assay

  11. Fluorescent focus assay

  12. Serotype specific quantification using RT-PCR for A. C. Dengue 1-strain TH-Sman Dengue 3- strain H87 D. B. Dengue 4-strain H241 Dengue 2- strain TH-36

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