Background: Escherichia coli (E. coli) is one of the most common agents of bacterial infections, the emergence of multidrug resistance in E. coli has posed major threat to global health resulting in failure of treatment that leads to increased mortality and cost of healthcare facilities. Material and Methods: For this study, a total of 500 specimens like urine, blood, pus, stool, sputum and different body fluids received in the Clinical Microbiology Laboratory were processed and inoculated on culture plates (Blood agar, MacConkey agar, Cysteine Lactose Electrolyte Deficient Agar) and incubated at 37°C for 18-24 hours. After identification, E. coli was processed for Antibiotic susceptibility testing on Muller Hinton Agar using Kirby Baeur disk diffusion method. E. coli ATCC 25922 was used as reference strain. Results: Of 500 specimens received in clinical laboratory from different age groups, 180 were culture positive, of which 88 were E. coli isolates. 48(54.5%) E. coli isolates were obtained from female patients while 40(45.5%) were from male patients. Isolation of E. coli was highest in Urine (80.68%) followed by Pus and other samples. E. coli were 100% resistant to Ampicillin followed by ceftriaxone (85.2%) and ciprofloxacin (63.6%) whereas least resistance was observed in imipenem (21.6%), piperacillin-tazobactam (29.5%). Discussion: E. coli isolates exhibited high resistance to ampicillin &ceftriaxone. Consequently, an obligate need exists for antimicrobial resistance surveillance to provide clinically appropriate and cost effective therapy.