The structural and nonstructural recombinant proteins of HSV type 1 and type 2 were expressed in E. coli by fusion with cro-beta-galactosidase proteins. These proteins containing amino acid sequences of ICP4, ICP27, ICP47, ICP22, major capsid protein and major DNA-binding proteins of HSV types 1 and 2 reacted in immunoblotting with the corresponding anti-HSV sera from hyperimmune rabbits. The nonstructural recombinant proteins can be used for enzyme immunoassay detection of HSV1 or HSV2 type-specific antibodies in sera from patients with acute HSV infection.