Post-stroke fatigue is still a relatively unexplored, often neglected condition. The fatigue often manifests as physical and mental lack of energy, and many patients mention fatigue as one of the most difficult sequels to which they have to adjust. Our aim was to assess the fatigue in 35 three-month post-stroke patients and its relation to personal and stroke-related factors, and to compare them with 35 age- and sex-matched healthy control subjects. The main outcome measures for the fatigue were Fatigue Severity Scale (FSS) and Multidimensional Fatigue Inventory (MFI-20). Barthel index was used for functional status assessment, while anxiety and depressive symptoms were evaluated using Hospital Anxiety and Depression Scale (HADS). Study results showed fatigue to be frequent (45%) and often severe. Post-stroke patients showed higher values of general fatigue dimensions, physical fatigue and reduced activity. The number of subjects reporting fatigue problems on the FSS and MFI-20 scales was significantly greater in the stroke group (16/35; 45%) than in the control group (4/35; 11%; chi2 = 12.5; P < 0.001). Considering associations between fatigue and Barthel index score, age, sex, HADS-A (anxiety) and HADS-D (depression) scores, statistically significant associations were recorded between fatigue and anxiety (P = 0.01) and depression symptoms (P = 0.02). Global fatigue showed positive correlation with Barthel index score too (P = 0.01). Multivariate analysis showed that anxiety and depression symptoms could predict post-stroke fatigue (adjusted R2 = 0.556; P < 0.05). Clinical trials and follow-up studies to find an effective treatment for patients with post-stroke fatigue are warranted.