Hospital acquired infections caused by Non-fermentative gram-negative bacteria (NFGNB) have features that are of specific concern, detection of which is helpful for prevention and for better quality of healthcare in hospital premises. Present study was conducted to find out the nosocomial infections caused by NFGNB and its sensitivity pattern. Present microbiological study was carried out at C. U. Shah Medical College. Clinical samples which were included in this study were in accordance with inclusion criteria laid down in the research protocol, which were obtained during January 2018 to June 2020.Samples were collected and processed with the help of automated identification and antimicrobial susceptibility testing instrument i.e. Vitek – 2 as per standard guidelines. Statistical analysis was done by Microsoft Excel. Total 385 were classified as NFGNB from 1000 confirmed isolates causing Hospital acquired infection (HAI). Respiratory site infection was most common type of HAI i.e. 48%. While most common isolate was Pseudomonas sp.i.e. 42% followed by Acinetobacter baumannii i.e. 31%. Medicine ward showed maximum isolates i.e. 33%, followed by Surgery ward i.e. 23%. Maximum NFGNB isolates encountered from sputum sample i.e. 25% while blood shows only 6%. Trimethoprim/sulfamethoxazole and ciprofloxacinwere disclosedmaximum resistant. This study reports 385NFGNB isolates causing HAI. Such alarming resistance among NFGNB spreading the threat in medical care and epidemiology as they act as nosocomial pathogen.  These isolated pathogens being the major cause of illness particularly in hospitalized patients.