Background: Burkholderia cepacia (formerly Pseudomonas) is a gram-negative bacillus that can remain viable in low-nutrient water and is typically found in soil and moist settings. It is one of the leading causes of sepsis in infants, and it is spread by human contact with contaminated medical devices and disinfectants. B. cepacia has emerged as a significant opportunistic pathogen in hospitalized and immunocompromised patients, colonizing the lungs of individuals with cystic fibrosis. Hospital outbreaks have been associated to infected faucets, nebulizers, disinfection solutions, multi-dose antibiotic vials, drinking water, distilled water, flowmeters, nasal sprays, and ultrasound gels. We describe our investigation and successful management of a nosocomial Burkholderia cepacia sepsis outbreak in a tertiary care hospital's newborn intensive care unit in Peshawar, Pakistan.Methodology: Blood samples from 50 newborns with sepsis were collected using a standardized approach and incubated using an automated blood culture system (BACT/Alert 3D and BACTEC 9050). Disk diffusion and the Minimum inhibitory concentration methods were used to test antimicrobial susceptibility. Gram staining was used to identify bacteria, and API (Analytical Profile Index) 20 NE was used to characterize them biochemically. Environmental and epidemiological investigations were also conduct to investigate the source and route of infection.Results: All of the 50 patients admitted in NICU (Neonatal Intensive Care Unit) were included in this study, and had lately-onset neonatal sepsis, with B. cepacia. During an epidemic in the NICU from 30th, April to 21st, June 2021, B. cepacia was isolated from over 45 blood samples. In total 45 neonates 17(35%) were female and 28 (65%) were male. Average age of neonates was 14.88 days. Antibiotic susceptibility testing was performed on Muller Hinton agar using Kirby Bauer's disc diffusion method and interpreted according to Clinical Laboratory Standard Institute (CLSI 2021) guidelines. As B. cepacia has intrinsic resistant to polymyxin class (colistin sulphate) of antibiotics, the susceptibility pattern of all isolates were almost similar i.e. showing resistance to tetracycline 100% (minocycline) and cephalosporin 3rd generation 100% (ceftazidime). The isolates were 100% sensitive to fluoroquinolones (levofloxacin), carbapenem 100% (meropenem), chloramphenicol 100% and sulfonamide 100% (co trimaxazole).Conclusion: In present study, the index case might have been exposed to infection due to an intravenous fluid utilized for fluid and electrolyte replacement for intravenous delivery were the source of the current nosocomial outbreak and physiological state of low immunity (preterm, low birth weight, and mechanical ventilation). The rest of the cases might have been exposed to this organism due to inadequate hand hygiene/improper cleaning and disinfection practices. Timely reporting and implementation of infection control measures can play a significant role in curtailing this outbreak.