Background: Nutrition services in hospitals are services provided to patients that are adjusted to clinical conditions, nutritional status, and body metabolism. Organizing foods that are not sanitary and hygienic can prolong treatment, even cause nosocomial infections. Prevention and control of nosocomial infections is to prevent the occurrence of microbial pathogens, including through the behavior or habits of food handlers in the hospital. Purpose: To determine the effect of personal hygiene knowledge towards Bacteriological Quantity of Nutritional Care Unit Workers‟ Hand at PKU Muhammadiyah Gamping Hospital. Method: This research is quantitative research using analytic observational method with cross sectional design on 13 Nutrition Installation officers of Muhammadiyah Gamping PKU Hospital. Independent variable in this research is knowledge of personal hygiene and the dependent variable is bacteriological quantity of nutritional care unit workers‟ hand that get it from questionnaire and hand swab. Results: The level of personal hygiene knowledge of the nutrition installation officers of PKU Muhammadiyah Gamping Hospital on high category is 76.90% and in the moderate category is 23.10%. The number of hand germs of nutritional installation officers of PKU Muhammadiyah Gamping Hospital are 61.50% of nutrition installation officers had a bad number of hand germs and as much as 38.50% of nutrition installation officers had good numbers of hand germs. The highest number of germs is 3,000 CFU / cm2 and the lowest is 50 CFU / cm2. Knowledge about personal hygiene does not affect the number of hand germs. Conclusion: The level of personal hygiene knowledge of the nutrition installation officers of PKU Muhammadiyah Hospital on good category. The category of number hand germs of nutritional installation officers of PKU Muhammadiyah Gamping Hospital is bad. There is no effect between personal hygiene knowledge and the number of hand germs of nutrition installation officers at PKU Muhammadiyah Gamping Hospital.