P300 spellers are common brain-computer interface (BCI) systems designed to transfer information between human brains and computers. In most P300 detections, the P300 signals are collected by averaging multiple electroencephalographic (EEG) changes to the same target stimuli, so the participants are obliged to endure multiple repeated stimuli. In this study, a spatial-temporal neural network (STNN) based on deep learning (DL) is proposed for P300 detection. It detects P300 signals by combining the outputs from a temporal unit and a spatial unit. The temporal unit is a flexible framework consisting of several temporal modules designed for analyzing brain potential changes in the time domain. The spatial unit combines one-dimensional convolutions (Conv1Ds) and linear layers to generalize P300 features from the space domain, and it can decode EEG signals recorded using different numbers of electrodes. Both amyotrophic lateral sclerosis (ALS) patients and healthy subjects can benefit from this study. In the withinsubject P300 detection and the cross-subject P300 detection, our approach gained higher performance with fewer repeated stimuli than other comparative approaches. Furthermore, we applied the proposed STNN in the P300 detection challenge of BCI Competition III. The accuracy score was 89% in the fifth round of repeated stimuli, outperforming the best result in the literature (accuracy = 80%) to the best of our knowledge. The results demonstrate that the proposed STNN performs well with limited stimuli and is robust enough for various P300 detections.INDEX TERMS P300 detection, spatial-temporal neural network (STNN), deep learning (DL).