Background & Objectives: Uropathogenic Escherichia coli (UPEC) harbors virulence properties which are usually absent in non-pathogenic E. coli. The study was undertaken to characterize the properties like hemolysin production, serotypes and multi drug resistance (MDR) in routinely administered antimicrobial drugs in the UPEC isolates. Design & Methods: Laboratory confirmed 105UPEC isolates, obtained by semi quantitative culture method from clinically symptomatic cases of urinary tract infections (UTIs) along with fecal isolates of E. coli from 50 healthy individuals were included as controls. All isolates were subjected to serotyping and haemolysin production on sheep blood agar. Antibiorgram was done using modified Kirby-Bauer’s disc diffusion method by the standard Clinical & Laboratory Standard Institute (CLSI) guidelines. Results: All 105 serotyped strains showed high prevalence to O25 followed by O120 and others. Total 25 different serotypes were observed. Hemolysin production observed in 14 (13.33%) of uroisolates and 3 (6%) of control strains. MDR was found in 47 (44.76%) isolates with preponderance to 3 classes: Ampicillin, Co-trimoxazole and Gentamycin. No correlation between serotypes, haemolysin production and MDR was found. Interpretation & Conclusion: Antimicrobial susceptibility profiles are necessary to be evaluated in India where a severe misuse of antibiotics at all levels can be seen. Stringent policies of antimicrobial use and infection control in all hospitals is a need of the hour. Detection of hemolysin production, serotyping and MDR in the isolates is reasonably easy and screening them in clinical microbiology laboratory of a tertiary care hospital is a worth consideration.