title: Analysis of ganciclovir-resistant human herpesvirus 6B clinical isolates using quenching probe PCR methodology.
abstract: Quenching probe PCR (QP-PCR) analysis was used to determine the frequency of ganciclovir (GCV) resistance among clinical isolates of human herpesvirus 6B (HHV-6B) obtained from patients with primary viral infection and viral reactivation. Forty-two HHV-6B clinical isolates were repeatedly recovered from 15 hematopoietic stem cell transplant (HSCT) recipients, and 20 isolates were recovered from 20 exanthem subitum (ES) patients. Of the 15 HSCT recipients, 9 received GCV during the observation period; however, none of the ES patients were treated with GCV. Two established laboratory strains, Z29 and HST, were used as standards in this study. Regions 1 and 2 of the U69 gene of all of the clinical isolates demonstrated the same melting temperature as regions 1 and 2 of the Z29 strain. For region 3, the melting temperatures of all clinical isolates fell between the melting temperature of the plasmid containing the A462D single nucleotide polymorphism (SNP) and the melting temperature of the Z29 strain, and the melting temperatures profiles of all clinical isolates were similar to the melting temperature profile of the Japanese HST strain. As expected, none of the 20 clinical isolates recovered from the ES patients and the 14 isolates recovered from the HSCT recipients who did not receive GCV treatment carried the six known SNPs associated with GCV resistance. Interestingly, these six SNPs were not detected in the 28 clinical isolates recovered from the 9 HSCT recipients who received GCV. Additional sequence analysis of the U69 gene from the 15 representative isolates from the 15 HSCT recipients identified other SNPs. These SNPs were identical to those identified in the HST strain. Therefore, the rate of emergence of GCV-resistant HHV-6B strains appears to be relatively low, even in HSCT recipients treated with GCV.
authors.affiliation: Department of Clinical Pharmacy, Fujita Health University Hospital, Aichi, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Hiramatsu
authors.affiliation: Department of Clinical Pharmacy, Fujita Health University Hospital, Aichi, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Suzuki
authors.affiliation: Department of Clinical Pharmacy, Fujita Health University Hospital, Aichi, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Yamada
authors.affiliation: Faculty of Clinical Engineering, Fujita Health University School of Health Sciences, Aichi, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Ihira
authors.affiliation: Department of Food Sciences and Nutrition, Mukogawa Women's University School of Human Environmental Sciences, Aichi, Hyogo, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Isegawa
authors.affiliation: Department of Pediatrics, Fujita Health University School of Medicine, Aichi, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Kawamura
authors.affiliation: Department of Pediatrics, Fujita Health University School of Medicine, Aichi, Japan.
authors.forename: Erina
authors.initials: E
authors.lastname: Matsuoka
authors.affiliation: Department of Pediatrics, Fujita Health University School of Medicine, Aichi, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Miura
authors.affiliation: Department of Pediatrics, Fujita Health University School of Medicine, Aichi, Japan tetsushi@fujita-hu.ac.jp.
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Yoshikawa
mesh_terms: D000293:Adolescent; D000328:Adult; D024882:Drug Resistance, Viral / Q000235:genetics; D005260:Female; D015774:Ganciclovir / Q000494:pharmacology*; D015654:Herpesvirus 6, Human / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic / Q000235:genetics; D055815:Young Adult
chemical_list: D015774:Ganciclovir