title: Real-time PCR determination of human herpesvirus 6 antiviral drug susceptibility.
abstract: A quantitative real-time PCR assay was developed to determine the antiviral drug susceptibility of human herpesvirus 6 (HHV-6). After short-term culture of the virus, HHV-6 isolates' susceptibility to the antiviral ganciclovir (GCV) was determined by measuring the HHV-6 variant B (HHV-6B) DNA levels in culture supernatants and infected cells using real-time PCR. A total of 12 well-characterized GCV-sensitive or -resistant strains and clinical isolates were used. This new assay with real-time PCR readout permitted the rapid (3 days), objective, and reproducible determination of HHV-6 drug susceptibilities with no need for stringent control of the initial multiplicity of infection. Furthermore, the real-time PCR assay results showed good correlation (rs=0.95) with those from the conventional TCID50 (50% tissue culture infecting dose) reduction assay (TRA). Thus, the real-time PCR assay described in this report was found to be a suitable quantitative method for determining the susceptibility of HHV-6 to antiviral drugs. It is faster and simpler than the TRA, and it is amenable to use in the routine diagnostic virology laboratory.
authors.affiliation: Department of Infectious Disease Control, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan. isegawa@bact.med.osaka-u.ac.jp
authors.forename: Yuji
authors.initials: Y
authors.lastname: Isegawa
authors.forename: Masaya
authors.initials: M
authors.lastname: Takemoto
authors.forename: Koichi
authors.initials: K
authors.lastname: Yamanishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ohshima
authors.forename: Nakaba
authors.initials: N
authors.lastname: Sugimoto
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000032:analysis; D015774:Ganciclovir / Q000494:pharmacology*; D015654:Herpesvirus 6, Human / Q000145:classification / Q000187:drug effects* / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; D015774:Ganciclovir