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Abstract Background The incidence of Post Stroke Depression (PSD) in the Rehabilitation Stage is high, which can bring serious physical and psychological disorders to patients. However, there is still a lack of targeted tools for screening PSD in the rehabilitation stage. Therefore, the aim of this study was to evaluate the factor structure and reliability of a measurement instrument to screen for PSD in the rehabilitation stage. Methods A cross-sectional study was conducted on 780 hospitalized stroke patients who were within the rehabilitation stage from May to August 2020. Exploratory factor analysis (EFA) as well as first- and second-order confirmatory factor analysis (CFA) were performed to evaluate the factor structure of the newly developed Symptom Measurement of Post-Stroke Depression in the Rehabilitation Stage (SMPSD-RS). The reliability and validity of the SMPSD-RS were also verified using several statistical methods. Results EFA extracted a 24-item, five-factor (cognition, sleep, behavior, emotion, and obsession) model that can clinically explain the symptoms of PSD during the rehabilitation stage. A first-order CFA confirmed the EFA model with good model fit indices, and the second-order CFA further confirmed the five-factor structure model and showed acceptable model fit indices. Acceptable reliability and validity were also achieved by the corresponding indicators. Conclusion The SMPSD-RS was proven to have a stable factor structure and was confirmed to be reliable and valid for assessing PSD symptoms in stroke patients during the rehabilitation stage.