Background: To assess antimicrobial sensitivity profile of ESBL producing E. coli isolates from various clinical samples. Methodology: Seventy- Eight E. coli isolates recovered from samples including pus, urine, blood, cerebrospinal fluid (CSF), stool, sputum, and different body fluids from inpatient and outpatient department received in the bacteriology laboratory in the department of microbiology were selected. ESBL screening and confirmation along with antimicrobial susceptibility test was done by Kirby–Bauer disk diffusion method according to the Clinical Laboratory Standards Institute (CLSI) guidelines. Results: Out of 48 inpatients samples of E. coli, 11 were found in pus, 20 in urine, 10 in blood, 4 in stool and 3 in sputum. Out of 30 outpatient samples, 7 were found in pus, 13 in urine, 5 in blood, 3 in stool and 2 in sputum. The difference was significant (P< 0.05). Antimicrobial susceptibility pattern of E. coli in pus, urine and blood against Ampicillin was 34%, 27% and 35%. Against Piperacillin was 43%, 42% and 60%. Against Piperacillin/Tazobactam was 91%, 85% and 82.4%. Against Amoxicillin/Clavulanic acid was 82%, 70% and 72%. Against Cefoperazone/Sulbactam was 80%, 78% and 84%. Against Cefoperazone was 32%, 27% and 54%. Against Cefoxitin was 27%, 35% and 32%. Against Ceftriaxone was 26%, 25% and 31%. Against Aztreonam was 37% , 23% and 20%. Against Imipenem was 100%, 100% and 100%. Against Gentamicin was 76%, 49% and 66%. Against Amikacin was 81%, 62% and 45%. Against Ciprofloxacin was 55%, 52% and 48%. Against Ofloxacin was 52%, 56% and 53% respectively. There was 62% ESBL producer in inpatients and 47% outpatient samples. There was 38% non-ESBL producers in inpatient and 53% outpatient samples. A significant difference was observed (P< 0.05). There was significant difference in antimicrobial susceptibility pattern of ESBL producing E. coli in urine and blood against various antibiotics (P< 0.05). Conclusion: There was high prevalence of ESBL against inpatient and outpatient samples obtained from blood, pus and urine