Introduction: Intensive Care Units (ICU’s) provides a suitable environment for the proliferation and persistence of multidrug organisms. The burden of nosocomial infections among critically ill patients is much higher in developing countries. Aims and Objectives: The present study was conducted to know the prevalence of bacterial pathogens from various ICU’s of a tertiary care hospital and to determine their resistance profile. Materials and methods: A retrospective study was done on 188 isolates collected over a period of 10 months from January 2018 to October 2018. The isolates were identified by the conventional biochemical method and antimicrobial susceptibility testing was performed by Kirby bauer disc diffusion method. Results: The most common gram negative isolate obtained in this study was Klebsiella spp. About 90/188 (47.87%), followed by Pseudomonas aeruginosa 22/188 (11.70%), Acinetobacter spp. 21/188 (11.17%), and E. coli 20/188 (10.64%). The predominant gram positive cocci isolated were Enterococcus spp. 9/188 (4.79%). All gram negative bacilli were sensitive to Imipenem. About 23.33% of Klebsiella isolates were resistant to Piperacillin tazobactam and 18.89% to Cefoperazone sulbactam. The most effective drugs against gram negative bacilli were Imipenem, Piperacillin tazobactam, and Cefoperazone sulbactam. Conclusion: This study showed the prevalence of multidrug resistant Klebsiella spp. from various ICU’s. These isolates were resistant to third generation cephalosporins, quinolones, and aminoglycosides and were sensitive to carbapenems. Among the non-fermenting gram negative bacilli, the resistant rates were higher for Acinetobacter isolates compared to Pseudomonas aeruginosa.