Natural rubber latex (NRL) allergy has been recognized as a public health concern. NRL allergy frequently occurs in healthcare professionals. In this retrospective study we report the changing frequency of Type I NRL allergy amongst healthcare workers suffering from hand dermatitis referred to our department between 1996 and 2003. We identified 224 healthcare workers from the patch test database with a diagnosis of hand dermatitis who had undergone NRL skin prick testing (SPT). We report the SPT results, patch test results and diagnoses for each individual. The percentage of positive SPT to NRL in healthcare workers decreased from 62% in 1996 to 10% in 2003. Type IV allergy to fragrance mix (13%) was the most frequent cause of allergic contact dermatitis. Thiuram mix (8%) and carba mix (4%) were the most frequent indicators of type IV allergy to chemicals in rubber gloves. Fragrance allergy was the most frequent type IV allergen found in healthcare workers with hand dermatitis. We conclude that hand care preparations free from fragrance allergens should be available in all areas of clinical work.