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Introduction: Emergency Medical Services (EMS) is one of the vital links in the care chain, and its services need to be improved. These services can be available through mobile-based automation system, in which low usability level of these systems lead to decrease the acceptance, satisfaction, and confidence of users especially the emergency care team. The purpose of this study was the usability evaluation of a national mobile- based automation system among the pre-hospital emergency care team. Material and Methods: This cross-sectional study was conducted on prehospital emergency care team members in Semnan and Shahroud Universities of Medical Sciences in 2022. The usability evaluation of the mobile- based EMS automation system was done using the Software Usability Measurement Inventory (SUMI) questionnaire. Multiple logistic regression models were used to analyze data. Results: One hundred eighty-eight EMS team members from the 31 EMS centers in Semnan province participated in present study. The mean total usability score was 61.93±15.37, the highest mean score was related to the efficiency feature (67.19±19.85) and the lowest mean score was related to the learnability feature (48.21±29.29). There was a reverse and significant relationship between being a manager and the agreement with the usability (p=0.04, OR= -3.383, CI 95%=0.389-29549). Conclusion: This study showed that although an automation system may be widely used in a country, its usability could be at a low level. In order to improve the different function of these systems, it is suggested to participate various clinical experts include prehospital emergency care team in all stages of designing and developing these systems. [ABSTRACT FROM AUTHOR]