Nucleic acid hybridization, a method to determine effects of antiviral compounds on herpes simplex virus type 1 DNA synthesis

Abstract

An application of the nucleic acid hybridization technique to screen effects of antiherpes compounds on herpes simplex virus type 1 (HSV-1) DNA synthesis is described. Whole cells are applied to nitrocellulose filters, their DNA is denatured and fixed to the filter. The resulting DNA spots are hybridized to cloned nick-translated HSV-1 DNA and the amount of hybridization is monitored by autoradiography or scintillation counting. Six antiherpes compounds: bromovinyldeoxyuridine, acyclovir, (R)- and (S)-enantiomers of 9-(3,4-dihydroxybutyl)guanine, 9-(4-hydroxybutyl)guanine and forscarnet, were evaluated for their effects on HSV-1 DNA synthesis. The most active compounds were bromovinyldeoxyuridine and acyclovir, with mean 50% inhibition values (IC50) for four different HSV-1 strains of 0.3 microM and 0.8 microM, respectively. The (R)-enantiomer of the new antiherpes compound 9-(3,4-dihydroxybutyl)guanine was found to be more active than the (S)-enantiomer, with mean IC50s of 6.5 and 14 microM, respectively, while mean IC50s of 2.5 and 68 microM were obtained for 9-(4-hydroxybutyl)guanine and foscarnet, respectively.