Objectives: The purpose of current study was to identify particular procedures which are associated with a high risk of glove puncture and thereby assist in limiting the risk of infection, especially those in high risk group. Study Design: Observational study. Setting: Outpatient department of dentistry at Liaquat University hospital, Hyderabad. Period: August 2017 to January 2018. Materials and Methods: In present study gloves perforation were assessed after performing various dental treatments by using Latex Biogel gloves. Total 400 gloves were collected from department of oral and maxillofacial surgery, orthodontics, operative dentistry and prosthodontics, 100 from each department. Simple Water Inflation technique was used to test punctures in gloves to determine the number and position of punctures in used gloves. Data collection was done using SPSS version 22.0. Descriptive statistics were computed and differences between groups were assessed through T- test. P-value ≤ 0.05 was taken as statistically significant. Results: Total 400 gloves were examined 100 from each department. Out of 100, 94 (94%) were found perforated from oral & maxillofacial surgery, 90 (90%) from orthodontics, 73 (73%) from prosthodontics & 80 (80%) from operative dentistry. Gloves puncture of left hand were more 292 (73%) than that of right hand 108 (27%) out of 400 gloves. Conclusion: Perforation of Gloves while several dental procedures have been evaluated and resulted that gloves on left hand were at greater risk unambiguously in the procedures in which wires were used, additional precautions like wearing double gloves during these procedures may therefore be indicated.