Background: Food safety is an international major concern in all food establishments, such as grocery stores. Food retailers provide a significant service to the public. Food safety-related procedures, and practices grocery stores are critical in protecting public health. The Egyptian retail food industry is expanding in size and popularity. In 2021, the Egyptian food retail sector recorded sales of $31.3 billion. The knowledge, attitude, and practices of food handlers are the most important factors in ensuring food safety. However, there is a scarcity of studies conducted to evaluate these factors in Egypt. Aim: The current study aimed to evaluate the level of food safety knowledge of food handlers at food retail sector in Egypt. Methodology: A cross-sectional study was conducted among 262 food handlers in food retail stores in Egypt. Food handlers were investigated using a self-administrated questionnaire that covered three major topics concerning food safety knowledge. Results: A major part of food handlers 82.4% were men while women were 17.6%. About 64% of food handlers had adequate knowledge of food safety, almost one third of food handlers (36%) had poor knowledge scores, and the knowledge score was (69% and 49%) about cleaning and temperature control, respectively.