For setting up an Enzyme-Linked Immunosorbent Assay (ELISA) to detect antibodies to herpes simplex virus type 1 (HSV-1), the virus was propagated in Vero cells and partially purified by sonification and ultracentrifugation on 30% sucrose solution. DEAE ion-exchange column chromatography was used for purification of goat anti-human IgG serum. The anti-human IgG immune serum and alkaline phosphatase were conjugated by glutaraldehyde method. ELISA test was performed by reacting HSV-1 antigen coated in polystyrene tubes with serum specimens and enzyme-IgG conjugates. The color produced by enzyme-substrate reaction was measured on a spectrophotometer. The results obtained by the ELISA had a good agreement with those obtained by a standard neutralization procedure on 119 serum specimens tested for antibody to HSV-1.