Objective: Real-time intramuscular electromyogra-phy (iEMG) decomposition, which is needed in biofeedback studies and interfacing applications, is a complex procedure that involves identifying the motor neuron spike trains from a streaming iEMG recording. Methods: We have previously proposed a sequential decomposition algorithm based on a Hidden Markov Model of EMG, which used Bayesian filter to estimate unknown parameters of motor unit (MU) spike trains, as well as their action potentials (MUAPs). Here, we present a modification of this original model in order to achieve a real-time performance of the algorithm as well as a parallel computation implementation of the algorithm on Graphics Processing Unit (GPU). Specifically, the Kalman filter previously used to estimate the MUAPs, is replaced by a least-mean-square filter. Additionally, we introduce a number of heuristics that help to omit the most improbable decomposition scenarios while searching for the best solution. Then, a GPU-implementation of the proposed algorithm is presented. Results: Simulated iEMG signals containing up to 10 active MUs, as well as five experimental fine-wire iEMG signals acquired from the tibialis anterior muscle, were decomposed in real time. The accuracy of decompositions depended on the level of muscle activation, but in all cases exceeded 85%. Conclusion: The proposed method and implementation provide an accurate, real-time interface with spinal motor neurons. Significance: The presented real time implementation of the decomposition algorithm substantially broadens the domain of its application.