A double-antibody enzyme immunoassay was developed for the identification and typing of herpes simplex virus (HSV) by employing a polyclonal rabbit capture antiserum together with type-common and type 2-specific monoclonal antibodies as detectors. The test successfully identified 45 type I isolates and 30 type 2 isolates as HSVs. Compared with immunofluorescent staining and restriction endonuclease analysis, enzyme immunoassay correctly typed 45 type 1 and 30 type 2 HSV isolates. Enzyme immunoassay was 100% sensitive for identification of HSV as compared with cell culture and 100% specific for typing as compared with immunofluorescence and restriction endonuclease analysis. Electron microscopy analysis suggested that approximately 10(6) virus particles were required for the identification and typing of HSV by enzyme immunoassay.