Acinetobacter species are Gram negative nonfermentative bacteria that have now emerged as important nosocomial pathogens involved in outbreaks of hospital infections.They are considered as opportunistic pathogens that readily colonize patients with compromised host defenses especially in intensive care units (ICUs), neonatal units, and surgical wards. The current study was conducted to type the Acinetobacter isolates obtained from various sources by a simplified phenotypic identification scheme and also to determine their antimicrobial susceptibility. Specimens like blood, CSF, endotracheal aspirate, urine, sputum, pus, bronchoalveolar lavage, HVS and body fluids were processed by standard methods and the antibiotic-sensitivity was performed by Kirby-Bauer disk diffusion technique as per Clinical and Laboratory Standards Institute guidelines (CLSI). The study was conducted in a tertiary care hospital for a period of 6 months (July 2013 – Dec 2013) in which out of a total of 1272 culture positive specimens, 53 Acinetobacter isolates were obtained from various specimens. Speciation was done in which predominance of A. baumannii( 90.6%) was seen while A. lwoffii and A. haemolyticus showed an isolation rate of 5.7% and 3.8% respectively. High levels of resistance were seen for Ampicillin –Sulbactam (96%), Ampicillin (94%), Aztreonam (94%), Cefuroxime (92%), ceftazidime (91%). The p-value was found to be statistically significant for all the above mentioned antibiotics except for Polymyxin B for which 100% sensitivity was recorded. Clinical co-relation must be under taken to exclude commensal contaminants, before considering it to be a pathogen and prescribing antibiotics to the patient.