Several studies have shown that acute psychosocial stress and chronic psychosocial stress reduce heart rate variability (HRV). It is likely that individuals suffering from burnout have reduced HRV, as a consequence of the long-term stress exposure. This study investigated HRV in 54 patients with clinical burnout (40 women and 14 men) and in 55 individuals reporting low burnout scores (healthy; 24 women and 31 men) and 52 individuals reporting high burnout scores (non-clinical burnout; 33 women and 19 men). The participants underwent a 300s ECG recording in the supine position. Standard deviation of normal R-R intervals (SDNN) and the root mean square of successive normal interval differences (RMSSD) were derived from time domain HRV analysis. Frequency domain HRV measures; total power (TP), low frequency power (LF), high frequency power (HF), and LF/HF ratio were calculated. All HRV measures, except LF/HF ratio, were lower in the clinical burnout patients compared to both the non-clinical burnout group and the healthy group. The difference was larger between the patients and the healthy group than between the patients and the non-clinical burnout group. HRV did not differ significantly between the non-clinical burnout group and the healthy group. Low HRV in burnout patients may constitute one of the links to associated adverse health, since low HRV reflects low parasympathetic activity - and accordingly low anabolic/regenerative activity.