The objective of this study is to analyze the most associate factors to Escherichia coli bacteria contamination in food at faculty canteen of University X. The design of this study is cross sectional design, by using primary data through laboratory test of 70 food samples and direct interview to 70 food handlers with questionnaire. The laboratory test results showed that 60% food were contaminated with the Escherichia coli bacteria. There were significant association between hygiene and sanitation of cooking utensils (p=0.005) and canteen environment (p=0.010) with Escherichia coli food contamination and statistically no significant between hygiene and sanitation of food handlers, food processing and canteen facilities with Escherichia coli food contamination. Based on multivariate analysis, the most influenced factors in this research were hygiene and sanitation of food processing, cooking utensils and canteen environment. Therefore, training of food handlers are necessarily in order to minimize Escherichia coli food contamination.