The rise in antibiotic resistance has been a major source of public health concern. As a result, mortality and morbidity rates have risen significantly. This study was done to identify gram-negative organisms causing bacteremia/sepsis, study their prevalence rates, and antimicrobial resistance patterns, as evidence-based knowledge of gram-negative organisms causing sepsis and their resistance profiles is essential for effective hospital control and better management of infections caused by resistant bacteria. A retrospective study, conducted from January 2016 to December 2019, blood samples were collected using aseptic guidelines and cultured using automated blood culture methods. Biochemical tests were done according to microbiology standard procedures, while antimicrobial testing was done according to CLSI guidelines. A total of 13,808 blood samples were received within the study period of four years. Of the total, 2079 showed significant growth, with 765 being GNB isolates. The most common isolates were Escherichia coli (35.42%), Klebsiella pneumonia (19.74%), Acinetobacter species (9.67%), and other non-fermenting gram-negative bacilli (11.76%). Escherichia coli showed yearly resistance to aminoglycosides, cephalosporins, penicillin, fluoroquinolones, and B-lactam combination agents. Routine surveillance and awareness of the prevalence, etiological agents, and antibiotic resistance of gram-negative bacteria causing bacteremia/sepsis is critical for individual therapy, hospital control, and the effectiveness of preventive interventions.