Phenotypic and genotypic characterization of acyclovir-resistant clinical isolates of herpes simplex virus.
Sixteen herpes simplex virus type 1 (HSV-1) and four type 2 (HSV-2) isolates resistant to acyclovir (ACV) were characterized retrospectively for drug resistance. Phenotypic testing was performed by means of tetrazolium reduction assay and genotypic analysis was carried out by sequencing of thymidine kinase (TK) and DNA-polymerase (pol) genes. All strains were characterized as cross-resistant to penciclovir, brivudin and susceptible to cidofovir. In addition, three strains were resistant to foscarnet. Genotypic analysis revealed two to seven non-synonymous mutations in the TK gene of HSV-1 and one to seven non-synonymous mutations in the DNA pol gene of HSV-1 and 2 associated with the gene polymorphism. Seventeen strains contained at least one non-synonymous resistant-related mutation in the TK gene and three strains, which were additionally foscarnet-resistant, revealed one resistance-associated mutation in the DNA pol gene. In most strains, resistant-related mutations in TK gene represented frameshift mutations and single non-synonymous nucleotide substitutions of conserved gene regions. However, numerous amino acid changes could not be interpreted clearly as accounting for resistance. In conclusion, further studies, e.g. site-directed mutagenesis experiments are required to characterize mutations of the TK and DNA pol genes in ACV-resistant viral strains as part of viral gene polymorphism or as cause of drug resistance.
Sauerbrei
A
A
Institute of Virology and Antiviral Therapy, German Reference Laboratory for HSV and VZV, Friedrich-Schiller University of Jena, Hans-Knoell-Strasse 2, 07745 Jena, Germany. Andreas.Sauerbrei@med.uni-jena.de
Deinhardt
S
S
Zell
R
R
Wutzler
P
P
D000212:Acyclovir / Q000031:analogs & derivatives / Q000494:pharmacology*; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001973:Bromodeoxyuridine / Q000031:analogs & derivatives / Q000494:pharmacology; D002470:Cell Survival; D002478:Cells, Cultured; D000077404:Cidofovir; D003596:Cytosine / Q000031:analogs & derivatives / Q000494:pharmacology; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D024882:Drug Resistance, Viral; D005090:Exodeoxyribonucleases / Q000235:genetics; D017245:Foscarnet / Q000494:pharmacology; D006147:Guanine; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology*; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology*; D006801:Humans; D008826:Microbial Sensitivity Tests; D020125:Mutation, Missense; D063065:Organophosphonates / Q000494:pharmacology; D010084:Oxidation-Reduction; D017422:Sequence Analysis, DNA; D013194:Staining and Labeling / Q000379:methods; D013778:Tetrazolium Salts / Q000378:metabolism; D013937:Thymidine Kinase / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics
D000998:Antiviral Agents; D063065:Organophosphonates; D013778:Tetrazolium Salts; D014764:Viral Proteins; C020235:brivudine; C053539:penciclovir; D017245:Foscarnet; D006147:Guanine; D003596:Cytosine; D013937:Thymidine Kinase; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; C043417:DNA polymerase, Simplexvirus; D001973:Bromodeoxyuridine; D000077404:Cidofovir; D000212:Acyclovir