The objective of this study is to determine the association of biofilm formation, extended spectrum beta lactamase [ESBL] production and antibiotic susceptibility pattern among uropathogenic Escherichia coli [UPEC] isolates. The present study was conducted on 137 UPEC isolates (counts > 105cfu/ml) from UTI cases. In all these isolates biofilm formation was detected by microtitre plate [MTP] method, ESBL production by combined disc diffusion technique and antibiotic susceptibility testing [AST] by Kirby- Bauer disk diffusion method. The data was analysed by using Medcalc software package. Chi-square test was applied. Among 137 UPEC isolates, 48 (35%) were biofilm producers [BFP’s]and 89 (65%) were biofilm non producers [BFNP’s] by MTP method. Of the total number of137 isolates, the highest number of strains were susceptible to amikacin followed by gentamicin, nitrofurantoin, cefepime.Among 137 isolates, ESBL producers were 28 (75%) of which 21 (75%) were BFP’s also, which makes a total of 49 (35%) (ESBL+Biofilm) and BFNP’s were 7[25%]. ESBL non-producing E.coliwere109 (80%), of which 32 (29.3%) were BFP’s and 77 (70.6%) were BFNP’s. The ability of biofilm formation was significantly higher in ESBL producing strains than that of ESBL non-producing strains (p<0.05). The ability of biofilm formation was found to be higher among ESBL producing stains of E. coli. Higher resistance rate was noted among biofilm producers to almost all the antimicrobial agents except few than non-biofilm producers.