Rapid detection and identification of Escherichia coli (E. coli) is essential to prevent its quickly spread. In this study, a novel fluorescence probe based on ZnTe quantum dots (QDs) modified by mannose (MAN) had been prepared for the determination of E. coli. The results showed that the obtained QDs showed excellent selectivity toward E. coli, and presented a good linearity in range of 1.0 × 105∼1.0 × 108 CFU/mL. The optimum fluorescence intensity for detecting E. coli was found to be at pH 7.0 with a temperature of 25 °C and incubation time of 20 min. Under these optimum conditions, the detection limit of E. coli was 4.6 × 104 CFU/mL. The quenching was discussed to be a static quenching procedure, which was proved by the quenching efficiency of QDs decreased with the temperature increasing.