Using AgNO3 and NaCl as enhnacers, highly SERS active AgNRs/AgCl nanosol substrate were prepared, in which the hydrophobic AgCl molecules formed and absorbed on the AgNRs surface by mens of strong hydrophobic forces between AgNRs and AgCl. The Vitoria blue B molecular probes on the surfaces of AgNRs/AgCl displayed a strong SERS paek at 1 611 cm-1. VBB was used to dye the Escherichia coli (E. coli), which exhibited the SERS proprerty with a Raman peak at 1 611 cm-1. Under the optimal conditions, the SERS intensities were linear to E. coli concentration in the range of 5×106～3×109 cfu·mL-1, with a detection of 2×106 cfu·mL-1. The new SERS method was applied for detection of E. coli in real samples, with features such as simplicity, rapidity and sensitivity. The results were in agreement with that of plate-count method.