Introduction - For more than a century, nosocomial infections, often known as hospital-acquired illnesses (HAI), have been acknowledged as a serious issue. Non-fermentative gram negative bacteria (NFGNB) have been linked to human illness more and more in recent years. A. baumannii is the most commonly encountered NFGNB, and it is frequently linked to high morbidity and mortality, including infections like meningitis, respiratory tract infections, urinary tract infections, bacteremia, and skin and soft tissue infections, particularly in patients with severe device-associated illness. Method- From June 2022 to December 2024, this study was carried out at the Index Medical College Hospital & Research Centre in Indore, Madhya Pradesh. The study included non-repeating samples taken from various intensive care unit patients, which were then forwarded to the microbiology lab for sensitivity testing and culture identification. Included were all patients who had been in intensive care units for more than 48 hours. The study excluded patients with less than 48 hours and those not in the intensive care unit. Every sample underwent microbiological processing. In accordance with CLSI standards 2020, bacterial identification and antibiotic resistance patterns were carried out. PCR was also used to validate the presence of the intrinsic blaOXA-51like gene in A. baumannii. Result - Twenty-eight Acinetobacter baumannii were identified from 200 patients in this prospective research. A. baumannii infections were discovered in 14% of cases (28 acquired infections out of 200) based on microbiological and clinical correlation. 18 (64.2%) respiratory samples had the highest percentage of isolates, followed by blood 7 (25%), urine 2 (7.1%), and pus 1 (3%) samples. The majority of the antimicrobial drugs did not affect the isolates at all. According to assessments of the isolates' antibiotic susceptibility, penicillin and cephalosporin resistance were the most prevalent, followed by gentamicin resistance (83%), and imipenem resistance (79%). Every Acinetobacter that was isolated was vulnerable to polymyxins and tigecycline.It was determined that all 28 isolates of A. baumannii carried the blaoxa-51-like gene. Discussion- The current investigation highlights Acinetobacter baumannii's increasing clinical importance as a leading nosocomial infection pathogen, especially in intensive care units. When it came to correctly identifying A.baumannii in a variety of clinical specimens, such as respiratory, bloodstream, and wound infections, molecular detection techniques proved to be successful. The presence of A. baumannii was found to be strongly associated with multidrug resistance, particularly to carbapenems and cephalosporins, which presents significant treatment problems.