Background: Bacterial biofilms play an important role in urinary tract infections and is responsible for persistence infections and also the higher antimicrobial resistance is seen in biofilm forming uropathogen as compared to free floating bacteria. So the present study was undertaken with the aim to know the prevalence of biofilm formation and antimicrobial resistant pattern of biofilm producer and non-biofilm producing uropathogens. Materials & Methods: A total of 146 Gram negative bacilli and 62 S. aureus isolated from patients suspected UTIs were tested for biofilm formation and antimicrobial susceptibility testing by Kirby-Bauer disc diffusion method on Mueller Hinton agar as per CLSI guidelines. Result: Out of 208 isolates from urine, Biofilm formation was noted in 122(58.66%) and no biofilm formation in 86(41.35%).[Strong Biofilm formation in 76(36.54%) and weak biofilm formation in 46(22.12%)ï.In our study, we noted biofilm and non-biofilm forming microorganism showed mark difference in antimicrobial resistance pattern. In Staphylococcus aureus striking difference was noted to ciprofloxacin (100% versus 33.33%) and azithromycin (96%versus 41.67%). Isolates showed no resistance to linezolid. Whereas isolates of Pseudomonas aeruginosa to netilin (100% versus 42.86%).And in other Gram negative bacilli difference was noted to gentamicin (87.93% versus 13.43%) and norfloxacin (84.48% versus 37.31%) Conclusion: Biofilm forming isolates showed higher antimicrobial resistance as compared to non-biofilm producer. Thus, Uropathogen should be routinely screened for biofilm formation.