A rapid quantitative PCR-based assay for testing antiviral agents against human adenoviruses demonstrates type specific differences in ribavirin activity.
Human adenovirus (HAdV) infections are increasingly frequent and potentially fatal as a disseminated disease in highly immunocompromised patients. Determining the in vitro sensitivity of HAdV to antiviral agents is not an easy task because HAdV CPE reduction assays are difficult to interpret and may take more than 1 week. We developed a phenotypic assay for testing the antiviral activity during the first round of replication using HAdV DNA concentration as an objective readout within 30 h. After evaluating the assay with cidofovir, we focused on determining the antiviral of ribavirin against different HAdV serotypes because clinical response of HAdV infections towards ribavirin treatment varied considerably. Several HAdV prototypes (1, 2, 5, 11, 31, 34, 48) associated with disseminated infections and clinical isolates were tested. Predominating HAdV of species C were more sensitive to ribavirin (HAdV-2 and -5: EC(50)<10 microM, EC(99) 111 and 104 microM, respectively) than HAdV of other species, for example HAdV-31 (EC(50) 56 microM, EC(99)>500 microM). Differential ribavirin sensitivity of HAdV types may contribute to the variable outcome of ribavirin therapy. Rapid screening of antiviral agents with the rapid qPCR-based assay against a multitude of HAdV serotypes may also facilitate development of future antiviral agents.
Stock
Rüdiger
R
Institute for Virology, German National Reference Laboratory for Adenoviruses, Hannover Medical School, Germany.
Harste
Gabi
G
Madisch
Ijad
I
Heim
Albert
A
D000258:Adenovirus Infections, Human / Q000821:virology; D000260:Adenoviruses, Human / Q000145:classification / Q000187:drug effects* / Q000235:genetics; D000998:Antiviral Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012254:Ribavirin / Q000494:pharmacology*; D013045:Species Specificity
D000998:Antiviral Agents; D015342:DNA Probes; D004279:DNA, Viral; D012254:Ribavirin