Introduction: Emergence of antibiotic resistance and biofilm formation among the nosocomial pathogens is of serious concern due to recurrence and chronicity of infections. The production of the extended spectrum-β-lactamases and their spread among bacterial pathogens is a matter of concern with regard to the future of antimicrobial therapy. Antimicrobial resistance is an innate feature of bacterial biofilms. Biofilm formation is higher in ESBL producing and MDR strains. The present study was undertaken with the aim to determine antibiotic resistance pattern of the isolates, to find prevalence of ESBL among isolates and their association with biofilm production. Material and Methods: The study was carried out in the tertiary care hospital for the period of 1yr, from November 2014 to October 2015. A total of 100 isolates of P. aeruginosa from various clinical specimens were taken and identified using standard microbiological procedures. The isolates were subjected to antimicrobial susceptibility testing and ESBL production as per CLSI method. Biofilm formation was achieved according to the Tissue Culture Plate (TCP) assay. Results: Out of these 100 isolates, 58 were multi drug resistant and 20 were extreme drug resistant. Forty nine (49.0%) isolates were ESBL producers, while 51(51.0%) isolates were non-ESBL producers. Seventy-eight isolates produced biofilm, out of which, 43 were found in ESBL producers and 34 in non-ESBL producers. Conclusion: ESBL and biofilm producing nosocomial pathogens are of significant concern as they can cause chronic relapses and dissemination of drug resistant isolates. Both ESBL and biofilm production in the isolates further increase the morbidity and mortality in hospitalized patients. Hence it is necessary to detect the biofilm forming bacteria, conduct surveillance of the antibiotic susceptibility and device effective infection control measures.