Introduction: The safety culture of the patient is a contributing factor for the maintenance of the user’s well-being in the health system because, through it, an organized systematization and quality of patient care are obtained, preventing possible intercurrences that can cause damages. Objective: To analyze the Patient Safety Culture (PSC) from the perspective of health professionals at the Reference Hospital of the Upper Juruá River, in the Brazilian Western Amazon. Methods: This is a cross-sectional study developed in a medium-sized public hospital in a municipality in Western Amazonia. The Survey for Patient Safety Culture survey of the Agency for Healthcare Research and Quality was applied to 280 professionals from December 2016 to February 2017. Descriptive analysis of the data and the internal consistency of the instrument were performed. Results: The results indicate the best evaluations in the dimensions of Teamwork in the scopes of the units (60%) and Organizational learning (60%). The aspects with the worst results were the dimensions of non-punitive responses to errors (18%) and frequency of events reported (32%). The internal reliability (Cronbach’s Alpha) analysis of the dimensions ranged from 0.35 to 0.90. Conclusion: The "culture of fear" seems to predominate in this hospital, however, the study showed that there is scope for improvement in all dimensions of CSP. The values of Cronbach’s Alpha presented similarity to the results obtained by the validation process.