Study objectives To assess the patient safety culture in Primary Health Care (PHC) setting after the transition to the Family Health Strategy (FHS) model in a Brazilian metropolitan area and compare the results between the categories of health care professionals. Methods A cross-sectional study including 246 workers from primary health care services in Federal District, Brazil. Data collection took place from October to December 2019 through the Medical Office Survey on Patient Safety Culture (MOSPSC) application. Patient safety culture was considered positive when the score was above 60%. For comparisons between the categories of health care professionals’, the ANOVA and Kruskal-Wallis test were used for composite percent positive scores, and Pearson’s chi-square or Fishers exact test for frequency and percentage of positive responses. Results The overall MOSPSC composite percent positive score was 49.9%. Among the 12 dimensions, only three showed a positive patient safety culture: Teamwork (73.1%), Organizational learning (62.9%), and Patient care tracking/follow-up (60.1%). The percentage of positive responses on overall quality assessment (78.1%) and overall patient safety assessment (78.0%) showed a positive evaluation. There was no significant difference in the composite percent positive score of overall MOSPSC (p = 0.135) and the percentage of positive responses on overall patient safety assessment (p = 0.156) between the categories of health care professionals. Overall quality assessment showed a significant difference between job roles (p < 0.001), in which nursing /health care technicians showed a significantly lower score than other job roles. Conclusion The patient safety culture assessment showed a weakness in the patient safety in the PHC services. The MOSPSC and nine of its dimensions presented a negative safety culture assessment, regardless of the high scores in the overall patient safety and quality assessments.