Minas fresh cheese is susceptible to microbial contamination that may occur during or after processing, including cross-contamination. Escherichia coli is a microorganism that could be found in these foods causing various diseases. The diarrheagenic Escherichia coli has virulence factors and one of the most important is the enterohemorragic. Among them, E. coli O157:H7 is one of the most important. The presence of eventual pathogens may turn, in a few days, cheeses unacceptable for consumption. The aim of this study was to detect the occurrence of flicH7 and rbfbO157 genes from E. coli by PCR in strains isolated from processing chain of Minas fresh cheese in the São Paulo State, Brazil. Twelve visits were performed in six dairies processing cheese. In each visit were collected samples from raw milk, pasteurized milk, cheese and swabs from environment (equipment, utensils and surfaces). For E. coli isolation, multiple tube technique was performed, the samples were incubated in LST culture (35°C/24-48h) following incubation in EC culture for quantification of thermotolerant coliforms (45°C/24h-48h). Then, the positives tubes were inoculated in Macconkey agar and Eosin methylene blue agar. The typical colonies were used to perform biochemicals tests (citrate, indol, methyl red and voges proskauer). After, PCR to detect genes encoding virulence factors was performed .E. coli was detected in every dairy processing plant and a total of 115 samples was observed, theflicH7 and rbfbO157 genes occurring in 5 plants. The presence of E. coli flicH7 gene was observed in 28 samples and 7 samples of rbfbO157 genes and 1 isolated carried both. The strains were isolated from floor, milking machine, raw milk, table, cheese, clot, drain. The results showed that consumption of Minas fresh cheese represents a possible risk to consumers and the producers need to improve the hygienical condition of the processing chain.