In this report, the E. coli detection method was developed by using glycine coated cobalt ferrite nanoparticles (Gly/CoFe2O4 ) as a collection probe from the drinking water sample. The Gly/CoFe2O4 nanoparticles were used to capture and separate E. coli from the drinking water. The detection is based on beta-glucuronidase enzyme present on the bacteria lysate. The enzyme activity was measured using 4-methylumbelliferyl β-D-glucuronide hydrate as fluorogenic substrates. The calibration curve of the E. coli showed the linear range to be between 1 × 102 ~ 1 × 104 CFU/ml (R2 =0.9944). The minimum detectable concentration was 1 × 102 CFU/mL. The developed new method offered the best way to determine E. coli from the water sample with a total analysis time of 45 min.