A serological survey using antibody to Epstein-Barr virus (EBV)-specific DNase activity as a marker for the identification of patients with nasopharyngeal carcinoma (NPC) has been carried out on healthy subjects who visited Government Employees' Clinic Center (GECC) for routine health examination and on individuals residing in NPC high-risk areas (HRA) in Taiwan. During a 3-year prospective study, 22,596 and 9,869 sera were collected from the GECC and HRA groups, respectively. Taking neutralization of 2 or more units of EBV DNase activity as a positive response, the positivity rates in the GECC and HRA groups were 5.4% and 11.92%, respectively. Among the antibody-positive individuals, three cases of NPC were found in the GECC group (detection rate 0.63%) and 11 in the HRA group (detection rate 1.32%). A further patient at stage III of the disease was found in the first year of following up of 1,005 antibody-positive individuals. Among the 12 NPC patients in the HRA, five were newly diagnosed as having stage II (three patients) and stage III (two patients) NPC. These results support the hypothesis that antibody against EBV-specific DNase activity may be a useful marker for detection of patients with NPC, and they imply that individuals having high levels of antibody to EBV DNase activity may have an increased risk of development of NPC.