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Aims: To assess safety culture in primary healthcare centres and to explore its associated factors. Methods: This multicentre cross-sectional descriptive study was conducted in the 30 primary healthcare centres in central Tunisia and used the validated French version of the Hospital Survey on Patient Safety Culture questionnaire’ (HSOPSC). Results: A total of 214 professionals participated, corresponding to a response rate of 85%. The “Teamwork within centres” dimension had the highest score (71.5%). However, two safety dimensions had very low scores: “Frequency of event reporting”, “Non-punitive response to errors” with percentages of 31.4% and 35.4%, respectively. Among the associated factors, the “Frequency of reported events” dimension was significantly higher among professionals involved in risk management committees (p = 0.01). Conclusion: This study demonstrates that the level of safety culture needs to be improved in primary healthcare centres in Tunisia and also highlights the need to implement a quality management system in primary healthcare centres.