The objective of this study is to determine the prevalence of natural rubber latex allergy and its associated factors among health care workers in Jordan based on a self-administered questionnaire. In this cross-sectional study, the data were obtained through a self-reported hand-delivered questionnaire that was distributed to 500 health care workers who use natural rubber latex gloves. A total of 420 (84%) health care workers responded by filling the questionnaire. Of the total number of 420 health care workers who filled the questionnaire, 57 (13.6%) reported allergy to latex gloves. The prevalence of allergy to latex gloves was significantly higher for those who had allergy to some foodstuffs (especially avocado, kiwifruits and chestnuts) or with history of eczema and hay fever. However, allergy to latex gloves was independent of gender, age, job, family history, type of gloves, hours of use/day and number of years of use. It is concluded that a high rate of allergy to latex gloves was reported among health care workers in Jordan.