| Summary: | Fever of unknown origin (FUO) is a medical term describing fever that lasts for at least three weeks without a diagnosis being reached after extensive diagnostic evaluation. Therefore, this study aimed to identify the common pathogens causing FUO in patients admitted to Abbasia Fever Hospital in Egypt from January 2020 to December 2022, their antimicrobial susceptibility profiles, and associated resistance genes. The study also aimed to investigate the burden of multidrug-resistant (MDR) pathogens and the priority pathogens nominated by the World Health Organization (WHO) for posing the greatest threat to human health due to antibiotic resistance. During the study period, about 726 patients were diagnosed with FUO. After extensive investigations, the cause of the FUO was found to be infectious diseases in 479/726 patients (66.0%). Of them, 257 patients had positive bacterial cultures, including 202 Gram-negative isolates that comprised <i>Klebsiella pneumoniae</i> (85/202; 42.1%), <i>Escherichia coli</i> (71/202; 35.1%), <i>Acinetobacter baumannii</i> (26/202; 12.9%), and <i>Pseudomonas aeruginosa</i> (14/202; 6.9%) and 55 Gram-positive isolates, including <i>Staphylococcus aureus</i> (23/55; 41.8%), <i>Streptococcus pneumoniae</i> (7/55; 12.7%), and <i>Enterococcus</i> spp. (25/55; 45.5%). The MDR phenotype was shown by 68.3% and 65.5% of the Gram-negative and Gram-positive isolates, respectively. Carbapenem resistance (CR) was shown by 43.1% of the Gram-negative isolates. Of the 23 <i>S. aureus</i> isolates obtained from research participants, 15 (65.2%) were methicillin-resistant <i>S. aureus</i> (MRSA). A high-level aminoglycoside resistance (HLAR) phenotype was found in 52.0% of the <i>Enterococcus</i> sp. isolates. The PCR screening of resistance genes in the MDR isolates showed that <i>bla</i><sub>OXA−48</sub> was the most prevalent (84%) among the carbapenemase-coding genes, followed by <i>bla</i><sub>VIM</sub> (9%) and then <i>bla</i><sub>IMP</sub> (12%). The ESBL-coding genes <i>bla</i><sub>TEM</sub>, <i>bla</i><sub>CTX-M,</sub><i>aac(6′)-Ib</i>, and <i>bla</i><sub>SHV,</sub> were prevalent in 100%, 93.2%, 85,% and 53.4% of the MDR isolates, respectively. This study updates the range of bacteria that cause FUO and emphasizes the burden of multidrug resistance and priority infections in the region. The obtained data is of relevant medical importance for the implementation of evidence-based antimicrobial stewardship programs and tailoring existing empirical treatment guidelines.
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