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Patient safety culture is a vital component in ensuring high quality and safe patient care. Assessment of staff perception on existing hospital patient safety culture (PSC) is the first step to promote PSC. This paper is aimed to assess the patient safety culture in 1 big public autonomous general hospital in Hanoi, Vietnam. This cross-sectional study surveyed 638 healthcare professional utilizing the validated (Hospital Survey on Patient Safety Culture [HSOPSC]) in an online format. This study adhered to STROBE guidelines. The positive response rate was high, with a percentage of 74.2. The strongest areas are teamwork within units (91.3%) and organizational learning/continuous improvement (88.4%). The areas for improvement are staffing (49.4%) and non-punitive response to error (53.1%). Hospital administrators should strengthen the culture of patient safety by formulating strategies and implementing interventions with emphasis on adequate staffing and promoting blame-free working environment.