Work stress and burnout have become important issues. Changes in work patterns frequently, long working hours, and too much pressure among workers in high-tech companies may result in the chronic fatigue symptoms and increase the risk of cardiovascular diseases, even leading to sudden death. Changes of heart rate variability (HRV) can be treated as a warning from the autonomic nervous systems and as a long-term monitoring method for chronic disease, for example, cardiovascular diseases and sudden death. The purpose of this study was to explore the association between occupational burnout and HRV.Proportional stratified convenient sampling was performed and in total, 120 individuals participated in this study. Questionnaires and the "occupational burnout inventory" were used to collect biographical and burnout information. A novel wrist physiological monitor was used to measure autonomic nervous system-related data, including HRV, low-frequency (LF) %, high-frequency (HF) %, and LF/HF ratio. Descriptive statistics and logistic regressions were computed using SPSS 17.0 (Chinese version, IBM Corp., New York, NY).The results show that participants who were 20 to 25 years old, work overtime because of onerous personal tasks, had a medical history, and recently felt unwell have higher "personal burnout." "Participants with a graduate degree, with onerous tasks, who were indirect staff, and with a regular-hour job tended to overcommit to their work." Significant associations were found among medical history, recently feeling unwell, and "work-related burnout." There was a positive association between HRV and job seniority. LF%, HF%, and LF/HF ratio were significantly correlated with job category. "Work overcommitment" was related to LF/HF ratio among men. Some items in "personal burnout" and "work overcommitment" were also associated with HRV among women.The findings suggest that the measurement of HRV can be applied in occupational settings to assess burnout. It not only allows administrators to quickly select the colleagues who need health care, but also provides timely and appropriate care, thereby promoting the health of the worker.