title: Genotypic detection of acyclovir-resistant HSV-1: characterization of 67 ACV-sensitive and 14 ACV-resistant viruses.
abstract: Infections due to herpes simplex virus (HSV) resistant to acyclovir (ACV) represent an important clinical concern in immunocompromised patients. In order to switch promptly to an appropriate treatment, rapid viral susceptibility assays are required. We developed herein a genotyping analysis focusing on thymidine kinase gene (TK) mutations in order to detect acyclovir-resistant HSV in clinical specimens. A total of 85 HSV-1 positive specimens collected from 69 patients were analyzed. TK gene could be sequenced directly for 81 clinical specimens (95%) and 68 HSV-1 specimens could be characterized as sensitive or resistant by genotyping (84%). Genetic characterization of 67 susceptible HSV-1 specimens revealed 10 polymorphisms never previously described. Genetic characterization of 14 resistant HSV-1 revealed 12 HSV-1 with either TK gene additions/deletions (8 strains) or substitutions (4 strains) and 2 HSV-1 with no mutation in the TK gene. DNA polymerase gene was afterwards explored. With this rapid PCR-based assay, ACV-resistant HSV could be detected directly in clinical specimens within 24 h.
authors.affiliation: Laboratoire de Virologie, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, Bron Cedex, Lyon, France. efrobert@sante.univ-lyon1.fr
authors.forename: Emilie
authors.initials: E
authors.lastname: Frobert
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Cortay
authors.forename: Tadamasa
authors.initials: T
authors.lastname: Ooka
authors.forename: Fatiha
authors.initials: F
authors.lastname: Najioullah
authors.forename: Danielle
authors.initials: D
authors.lastname: Thouvenot
authors.forename: Bruno
authors.initials: B
authors.lastname: Lina
authors.forename: Florence
authors.initials: F
authors.lastname: Morfin
mesh_terms: D000212:Acyclovir / Q000494:pharmacology*; D000328:Adult; D002648:Child; D002675:Child, Preschool; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D024882:Drug Resistance, Viral; D005260:Female; D005838:Genotype; D006561:Herpes Simplex / Q000821:virology*; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods; D013937:Thymidine Kinase / Q000037:antagonists & inhibitors / Q000235:genetics; D014764:Viral Proteins / Q000037:antagonists & inhibitors / Q000235:genetics
chemical_list: D014764:Viral Proteins; D013937:Thymidine Kinase; D004259:DNA-Directed DNA Polymerase; D000212:Acyclovir