Maintaining proper sanitation and behavior of food handlers is crucial in determining the quality of food. In addition, the presence of Escherichia coli indicates food contamination, which can lead to foodborne illness. Therefore, this study aimed to analyze the role of sanitation hygiene and food handlers’ behavior on the occurrence of Escherichia coli contamination in food served to children at Elementary Schools and Madrasah Ibtidaiyah (MI). In this study, an analytical survey method with a cross-sectional design was employed. The population consisted of 78 individuals involved in selling children’s food during the January 2023 period. The sample consisted of 43 food vendors who were selected using a purposive sampling. The inclusion criteria were respondents who sell food in Elementary Schools and MI, and the exclusion criteria were those who sell covered or packaged food. Data were collected through observation sheets and interviews, and data analysis involved both univariate and bivariate methods, using the Chi-Square test. The results showed that 31 respondents (72.1%) adhered to sanitary hygiene requirements, 33 respondents (76.7%) fulfilled the criteria for food handlers’ behavior, and 9 respondents (20.9%) had their food contaminated with Escherichia coli. There is a relationship between sanitation hygiene and Escherichia coli contamination in children’s food in Elementary Schools and MI (p-value = 0.00). Additionally, a correlation was observed between food handlers’ behavior and Escherichia coli contamination in food served at both types of schools (p-value = 0.00). These results emphasized the importance of street food vendors consistently prioritizing and implementing proper food sanitation, hygiene and healthy practices. This method was crucial to prevent Escherichia coli contamination and ensure the safety of the food served to children.