The present invention relates to nucleic acid probes specific to E. coli, which is useful for detecting and identifying E. coli in a biological sample. More particularly, the present invention relates to a DNA chip for detecting and identifying E. coli, on which nucleic acid probes derived from 23 S rRNA gene of E. coli are immobilized. The application of the DNA chip according to the present invention allows time-saving and accurate diagnosis of bacterial infection compared with the conventional of bacterial culture methods. In addition, it is not affected by antibiotics addition in clinical practice, thus leading to more accurate diagnosis.