Objective: To evaluate the knowledge, attitude, and practices about personal protective equipment (PPE) among dental surgery assistants in a Public Sector Teaching Hospital of Multan. Methods: A cross-sectional survey was carried out among dental surgery assistants in the dental outpatient departments of a public sector hospital of Multan city from Aug to Oct 2019. Pre-structured questionnaire which comprised of socio-demographic data, questions related to knowledge, attitude and practices was used. In the domain of knowledge there were 6 items. Dental assistant who scores ≤4 were considered to have appropriate knowledge for PPE. In the domain of attitude there were 5 items. Dental assistant who scores ≤3 were considered to have positive attitude for PPE. In the domain of practices there were 6 items. Dental assistant who scores ≤4 was considered good practicing of PPE. Results: Of 80 participants, 65 (81.25%) were males and 15 (18.75%) were females. Majority (n=54, 67.5%) of them were between 21-30 years of age. Appropriate knowledge was observed in 77 (96.3%) participants, positive attitude in 53 (66.3%) while, only 8 (10%) participants practiced good PPE. A significant difference was reported in the practices of dental assistants when compared with their knowledge (p-value 0.046) while attitude was found to be insignificant (p-value 0.710). Conclusion: Dental surgery assistants of the renowned public sector teaching hospital of Multan city reported sound and apposite knowledge, moderate attitude while inappropriate practices about PPE.