Introduction: Dental clinics pose multiple ways of cross contamination. Personal protective equipment (PPE) is one of the effective ways to control the spread of infections among dentists, providers of oral care, health care professionals, and medical professionals. The aim of this study is to check the microbial load on reusable PPE suits before and after clinic duty in a dental college using ATP bioluminometer. Materials and Methods: The PPE sleeves of the dominant hand of 40 dental professionals were assessed at rest in two different time intervals in a given day. The first samples were collected before clinic duty began and the second sample was collected after clinic duty was completed. The samples collected and assessed using ATP bioluminescence test. Results: Among the 40 samples taken, all showed an increase in the microbial load after clinic duty. The increase is not constant among all subjects. The samples were collected only from the dominant hand sleeve before and after. The paired t test was done with the results obtained from this study. The value of P=0.000045; P is < .00001. The result is significant at P< .05. There was a significant difference in the microbial load on the PPE suits before and after clinic duty. Conclusion:The results of the study stresses the importance of wearing good and effective PPE suits while performing clinical procedures in order to reduce incidence of nosocomial infections.