Typing of Herpes Simplex Virus with Synthetic DNA Probes

Abstract

Three single-stranded oligonucleotide probes, 22 bases long, homologous to unique regions of herpes simplex virus (HSV) types 1 (HSV-1) and 2 (HSV-2) and a region common to both were chemically synthesized with use of a modified phosphochloridite protocol. For hybridization experiments each probe was labeled with use of polynucleotide kinase and [γ-³²P] ATP to a specific activity of ∼2 × 10⁹ cpm/μg. Two hundred one clinical isolates of HSV (96 HSV-1 and 105 HSV-2) collected from vesicles in the mucocutaneous junction of the mouth or from the genital area were analyzed. There was a 99% (199 of 201) agreement between hybridization and monoclonal antibody typing; the two discrepant isolates of HSV-2 that were negative by monoclonal antibody typing were confirmed as HSV-2 by restriction endonuclease analysis. The probes detected between 10⁴ and 10⁵ HSV infectious units and from 150 to 600 HSV-infected Vero cells. No binding was detected between any ofthe three probes and isolates of cytomegalovirus, Epstein-Barr virus, and varicellazoster virus.