© 2021. All Rights Reserved.Objective: Hand washing is not only the simplest universal method to prevent healthcare related infections, it is also very important for the protection and development of general public health. In this study, it was aimed to investigate the hand washing habits and load of microorganisms in the hands of healthcare professionals and non-healthcare participants, so to raise the awareness in terms of hand hygiene. Methods: The study group consisted of patients and their relatives over 18 years of age, medical staff and medical school students. A questionnaire consisting of 17 questions regarding their personal characteristics and hand washing habits were applied to all participants. Swab samples were taken by sterile swabs from 1 cm2 areas of four different regions of the hands that were predominantly used by the participants, and were inoculated on blood agar plates and evaluated after 48 hours of incubation. Results: A total of 194 participants were reached, including 77 patients and patient relatives, 85 medical staff and 32 medical school students. A hundred and thirty-five of the participants were women and 59 were men. Bacteria that are members of normal skin flora (coagulase negative staphylococci, viridans streptococci, coryneform bacteria, micrococci) were isolated from almost all samples and more than 100 CFU bacterial load was detected in 53 participants. In 59 (30.4%) participants, microorganisms not included in normal skin flora (Gram negative enteric bacteria, Bacillus spp., Pseudomonas spp., Enterococcus spp., Staphylococcus aureus, mold and yeast fungi) were isolated. The proportion of these was calculated as 23% in patient/patient relatives, 33% in medical staff and 41% in students (p> 0.05). Women had a significantly lower bacterial load than men, 40-49 age group had a significantly lower bacterial load than participants over 60, and intensive care unit staff had a significantly lower bacterial load than patient/patient relatives and medical staff other than intensive care unit. Some hand washing habits of the participants were evaluated by scoring; 2 points before eating, 1 after eating, 1 before restroom, 2 after restroom, 2 after money contact, 2 when entering home from outside. As the hand washing score increased, the bacterial load on the hands decreased, but it was not statistically significant. It was observed that factors such as smoking, long nails, liquid/solid soap, paper/fabric towel, having a pet did not affect the microorganism load on the hands. The presence of any wound/lesion, prolonged time after hand washing and use of moisturizer were found associated with increased load. Conclusion: Being aware of the importance of hand hygiene in the community and especially among healthcare professionals is extremely important for the correct application of hand washing practices. In this study, it was aimed to emphasize the importance of hand washing by demonstrating the microorganism load on hands with the hand washing habits in a group of hospital related participants.