Herpes Simplex Virus Isolates from an Immunocompromised Patient who Failed to Respond to Acyclovir Treatment Express Thymidine Kinase with Altered Substrate Specificity

Ten sequential post-treatment herpes simplex virus type 1 (HSV-1) isolates were obtained from an immunocompromised patient whose infection, during prolonged treatment, became unresponsive to acyclovir (ACV). Of the ten isolates, eight later isolates were resistant in vitro to ACV and ganciclovir (DHPG), but remained sensitive to 9-β-D-arabinofuranosyladenine (ara-A) and phosphonoformate (PFA). Biochemical characterization of plaque-purified clones of the resistant isolates revealed an altered thymidine kinase (TK) substrate specificity phenotype. The comparative nucleotide sequence analysis of polymerase chain reaction (PCR)-amplified DNA encoding the TK genes of one sensitive and two resistant clones showed a single mutation at nucleotide 527. This change would result in a substitution of arginine by glutamine at residue 176 of the polypeptide, a mutation previously observed in a laboratory isolated variant, SC16 Tr7 (Darby et al., 1986).