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This knowledge management (KM) study focuses on factors affecting KM at a Ministry of Public Health institute. The developed questionnaires were distributed to all institute staff. Four major factors were investigated in the survey: Organizational Culture, Information and Technology, KM Content, and Administration and Management. Statistical analysis was used to calculate the relationships among factors and KM according to the KM model from the Thai Knowledge Management Institute. There was a 78.8% response rate. Organizational Culture, Information and Technology, and Administration and Management were found to be significant factors associated with KM. In addition, other elements within these factors had a significant relationship with KM. However, KM content, organizational structure, and conflict did not have a significant relationship with KM. The authors found quality management systems i.e. ISO 9001, ISO 17025 or ISO 15189 played an important role in KM at the institute. The authors also found drawbacks and room for KM improvement in the institute. The results in this paper can be used as knowledge-based evidence to enhance and improve the institute's routine operation and its performance.