Catheter associated urinary tract infection's (CAUTI) account for more than 80% of all healthcare associated infections (HAIs) as compared to non-catheterized urinary tract infections.Catheter associated urinary tract infections occur on the third day after insertion of catheter in patients having urinary tract infection (UTI). In long term catheter use, bio-film form along the catheter which increases the risk of antibiotic resistant pathogens. Most common pathogens involved were Escherichia coli and Klebsiella pneumoniae which produce the enzymes Extended Spectrum β-Lactamases (ESBLs).Objective: To compare the frequency of ESBLs in catheterized and non-catheterized UTI infections. . Urine samples were cultured according to WHO protocol and antimicrobial Susceptibility testing was performed by Modified Kirby-Bauer disc diffusion Method. Escherichia coli and Klebsiella pneumoniae were tested for ESBL production by phenotypic confirmatory method of disk diffusion synergy using a disc of amoxicillinclavulonate (30μg) and ceftrixone (30μg), cefotaxime (30μg) and aztreonam (30μg) discs. Results: Out of 300 positive urinary isolates of Escherichia coli and Klebsiella pneumoniae from CAUTI, 65.33% were ESBL producing isolates whereas out of 300 positive urinary isolates of Escherichia coli and Klebsiella pneumoniae from non-catheterized UTI, 47.66% were ESBL producing isolates. The results were highly significant (p < 0.001). Conclusion: Results showed that frequency of ESBLs were higher in catheterized patients as compared to non-catheterized patients. This is suggestive of a need for regular screening and surveillance for ESBL producing organisms. Patients infected with these organisms should be nursed with contact precautions to avoid the spread of nosocomial infection.100% after 30 days [4]. Bacterial colonies form into the epithelial cells of the urinary tract which are responsible for the infection [5]. The infection that occurs with an indwelling Foley's urinary catheter within 48 hours is a catheter-associated urinary tract infection [6]. Indwelling Foley's urinary catheter enables emptying of the bladder. Urinary catheterization is categorized as short term that is in situ less than 28 days or long term is in situ more than 28 days [7]. The presence of bacteria in urine is known as bacteriuria which is either asymptomatic (CA-ASB) or symptomatic bacteriuria. Asymptomatic bacteriuria is usually present in CAUTI [8]. Patients with urinary tract infections have symptoms of fever, urinary urgency, lower abdominal pain, dysuria, pyuria, and leukocytes in urine [9]. The presence of bacteria with a count of 10 5 CFUs per ml is an indication of urinary tract infection [10]. The level of bacteria in urine increases from 10 5 to 10 8 CFU per ml within 24 to 48 hours if antimicrobial therapy is not given to the patient [11]. Bio-film formation along the catheter reflects the presence of organisms that increase resistance to the host immune response and to antibiotics. Bio-film forms when a catheter was inserted greater t...