Application of real-time PCR for determination of antiviral drug susceptibility of herpes simplex virus.
A quantitative real-time PCR (TaqMan) assay was developed for determination of antiviral drug susceptibility of herpes simplex virus (HSV). After short-time culture of the virus, the antiviral drug susceptibility of HSV isolates for acyclovir (ACV) was determined by measuring the reduction of the HSV type 1 (HSV-1) DNA levels in culture supernatants using real-time PCR. The 50% inhibitory concentration was reported as the concentration of antiviral drug that reduced the number of HSV-1 DNA copies by 50%. A total of 15 well-characterized ACV-sensitive or -resistant strains and clinical isolates were used for assay evaluation. The new assay with real-time PCR readout permitted rapid (3 days), objective, and reproducible determination of HSV-1 drug susceptibilities with no need for stringent control of initial multiplicity of infection. Furthermore, the real-time PCR assay results showed good correlation (r = 0.86) with those for the plaque reduction assay. In conclusion, the real-time PCR assay described here is a suitable quantitative method for determination of antiviral susceptibility of HSV-1, amenable for use in the routine diagnostic virology laboratory.
Department of Virology, Eijkman-Winkler Center, University Medical Center Utrecht, The Netherlands. r.stranska@lab.azu.nl
Růzena
R
Stránská
Anton M
AM
van Loon
Merjo
M
Polman
Rob
R
Schuurman
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002522:Chlorocebus aethiops; D003199:Computer Systems; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis; D018259:Herpesvirus 1, Human / Q000187:drug effects; D007700:Kinetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D018139:Simplexvirus / Q000187:drug effects*; D014709:Vero Cells; D010948:Viral Plaque Assay
D000998:Antiviral Agents; D004279:DNA, Viral