An enhanced neutralization assay was developed to permit the sensitive, specific, and reproducible measurement of antibodies to varicella-zoster virus (VZV). Optimal neutralization was achieved using a combination of guinea pig complement (C') and rabbit anti-human IgG. This provided 625-, 160- and 13- to 64-fold increases in dilution endpoints of human post-zoster serum, varicella-zoster immune globulin and representative sera from recipients of live attenuated varicella vaccine, respectively, above those measured in the absence of C' and anti-IgG. The specificity of the assay was shown by the absorption of serum neutralization capacity with VZV-specific antigen and the lack of concordance between antibody titers to VZV with those to either herpes simplex virus type-2 or cytomegalovirus. The antibody status of recipients of live attenuated varicella vaccine was established from the amount of neutralizing activity produced at a single optimal serum dilution.