The purpose of this prospective study is to determine the frequency and site of glove perforation during cardiac surgical procedures. Over a period of six months, gloves from 206 surgical team members were collected at the conclusion of surgery. All cases of known perforations were eliminated from the study. The percentage of glove perforation was 14%. The distribution of perforation across locations of the hand was significantly unequal (P = 0.001). We found that 73% of the punctures occurred in one of four contiguous locations on the glove: the radial side of the index finger (28%), the radial side of the thumb (21%), the palmar side of the index (14%), and the palmar side of the thumb (10%). Therefore, we recommend glove reinforcement on these locations that would provide better protection against transmission of infectious agents. Discomfort from restricted dexterity and impaired sense of touch with double gloving renders glove reinforcement a suitable alternative for universal precautions, especially in cardiac surgery while high level of perfection and dexterity were needed in lengthy, critical operations.