Introduction: To minimize the risk of cross-infection between the patient and the medical staff, it is necessary to use individual protective measures such as gloves. According to the recommendations of the Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO), protective gloves should always be used upon contact with blood, mucosa, injured skin or other potentially infectious material. Materials and Methods: The aim of the study was to evaluate, through quasi-observation, the use of protective gloves by medical staff according to the guidelines issued by the CDC and WHO. The results were subject to statistical analysis (p < 0.05). Results: During 1544 hours of observations, 3498 situations were recorded in which wearing protective gloves is demanded from the medical staff. The overall percentage of the observance of using gloves was 50%. The use of gloves depended significantly on the type of ward, profession, performed activity, number of situations that require wearing gloves during the observation unit and the real workload. During the entire study, as many as 718 contacts with patients were observed in which the same gloves were used several times. Conclusion: Wearing disposable protective gloves by the medical staff is insufficient.