Exposure to natural rubber latex may cause immediate hypersensitivity reactions. Published latex sensitization prevalence rates range from 2.9% to 22% among health care workers, and from 0.12% to about 20% of occupationally unexposed populations. In this study, self-administered questionnaires addressed job and personal characteristics, glove use, and symptoms in two groups of hospital workers: those who regularly used latex gloves and those who did not. Serum was tested for latex-specific immunoglobulin E. Air, surface, and air-filter dust samples for natural rubber latex were collected. The prevalence of latex sensitization was 6.3% in the non-users and 6.1% in the latex glove users (P = 0.9); 81.3% of sensitized workers were atopic compared with 59.5% of non-sensitized workers (P < 0.05). Reporting of work-related hand dermatitis was more common in the latex glove users (23.4%) than in the non-users (4.9%), as were rhino-conjunctivitis (16.3% and 7.9%, respectively, [P < 0.01]), and hand urticaria (9.9% and 2.1%, respectively, [P < 0.01]). There was no significant difference in work-related symptoms between the sensitized and non-sensitized workers. Environmental concentrations of latex were higher in the work areas of the non-sensitized workers, but higher in the clinical than in the non-clinical areas. Occupational latex glove use was not a risk factor for sensitization.