Background: Exposure to natural rubber latex, primarily through the use of gloves, is a well-recognized cause of occupational asthma. We investigated latex glove use among Australian workers and estimated the resultant burden of occupational asthma among healthcare workers (HCWs). Methods: Data were collected in 2014 as part of the Australian Work Exposures Study-Asthma, a telephone survey investigating the prevalence of current occupational exposure to asthmagens, including latex. We estimated adjusted prevalence ratios (aPRs) to determine variables associated with the use of latex gloves among HCWs and calculated the asthma-related disability-adjusted life years due to latex exposure among HCWs. Results: Latex gloves were used by 22% of respondents. Almost two-thirds (63%) of HCWs reported wearing latex gloves, with 26% using powdered latex gloves. The use of latex gloves was more common among those employed in micro companies (less than 5 employees) than large companies (200+ employees) (aPR = 1.5, 95% confidence interval 1.1-2.0). Latex exposure in HCWs was estimated to contribute 3% of the total asthma-related burden. Discussion: Latex gloves are widely used by Australian workers and by HCWs in particular. Conclusions: This is the first estimate of the burden of asthma attributable to occupational exposure to latex among HCWs. These results can be used to guide decisions regarding the control of occupational exposure to latex.