BACKGROUND: This study aimed to determine the prevalence of extended-spectrum-beta-lactamase (ESBL) and carbapenem-resistant gram-negative bacteria (GNB) isolated from patients at the King Faisal Hospital in Makkah, Saudi Arabia.METHODS: In this cross-sectional study, a total of 298 patients admitted to the intensive care unit for 48 hours and who had a central venous catheter were selected using a census sampling method. Only patients with ESBL and carbapenem-resistant GNB-isolated organisms (175 patients) were included. The susceptibility test of GNB was carried out according to the standard recommendations. The identified strains were tested in-vitro against several antimicrobial drugs. Statistical analysis was performed using SPSS version 24.RESULTS: 36(20.6%) of samples were ESBL-producing GNB, whereas 139(79.4%) were carbapenem-resistant GNB. The pooled proportional estimates of ESBL-producing GNB Escherichia coli, Klebsiella pneumoniae, and other GNB were 44.4%, 41.6%, and 14.0%, respectively; the pooled proportional estimates of carbapenem resistance GNB Klebsiella pneumoniae, Acinetobacter baumannii complex/hemolyticus and other GNB were 82.8%, 10.8%, and 6.4%, respectively. All ESBL-producing GNB and carbapenem-resistance GNB were multidrug-resistant pathogens. The highest carbapenem resistance GNB 139(100%) was to ampicillin, and the lowest 122(87.7%) was to Amoxicillin/clavulanic acid (Amox/clav). All ESBL-producing GNB 36 (100%) were resistant to cefotaxime, and 35 (97.2%) were resistant to ampicillin, cefuroxime, cefepime, and ceftazidime. Additionally, the effective antibiotic against ESBL-producing GNB was imipenem.CONCLUSION: Antibiotic utilization measures appear to contribute to the control of the emergence of multidrug-resistant pathogens such as ESBL and carbapenem-resistant GNB. Strict adherence to well-accepted infection control guidelines along with caution in using broad-spectrum antimicrobial agents represents the best strategy for preventing the emergence and spread of multidrug-resistant pathogens.KEYWORDS: Carbapenem, Extended-spectrum beta-lactamase, Gram-negative bacteria, Resistant, Saudi Arabia