Several studies have shown the positive clinical effect of brain computer interface (BCI) training for stroke rehabilitation. This study investigated the efficacy of the sensorimotor rhythm (SMR)-based BCI with audio-cue, motor observation and multisensory feedback for post-stroke rehabilitation. Furthermore, we discussed the interaction between training intensity and training duration in BCI training. Twenty-four stroke patients with severe upper limb (UL) motor deficits were randomly assigned to two groups: 2-week SMR-BCI training combined with conventional treatment (BCI Group, BG, n = 12) and 2-week conventional treatment without SMR-BCI intervention (Control Group, CG, n = 12). Motor function was measured using clinical measurement scales, including Fugl-Meyer Assessment-Upper Extremities (FMA-UE; primary outcome measure), Wolf Motor Functional Test (WMFT), and Modified Barthel Index (MBI), at baseline (Week 0), post-intervention (Week 2), and follow-up week (Week 4). EEG data from patients allocated to the BG was recorded at Week 0 and Week 2 and quantified by mu suppression means event-related desynchronization (ERD) in mu rhythm (8–12 Hz). All functional assessment scores (FMA-UE, WMFT, and MBI) significantly improved at Week 2 for both groups (p < 0.05). The BG had significantly higher FMA-UE and WMFT improvement at Week 4 compared to the CG. The mu suppression of bilateral hemisphere both had a positive trend with the motor function scores at Week 2. This study proposes a new effective SMR-BCI system and demonstrates that the SMR-BCI training with audio-cue, motor observation and multisensory feedback, together with conventional therapy may promote long-lasting UL motor improvement.Clinical Trial Registration: [http://www.chictr.org.cn], identifier [ChiCTR2000041119].