Chronic bacterial prostatitis is a persistent infection of the prostate characterized by frequent relapses due to incomplete eradication of the causative organisms, with a negative impact on patient’s quality of life. This study aimed to determine the most common bacterial causative agents and their antibiotic sensitivity patterns in patients with chronic bacterial prostatitis in Duhok, Kurdistan, northern Iraq. A standardMeares-Stamey four-glass test was performed for all males presenting with chronic prostatitis symptoms for more than 3 months. Men with high leukocyte counts and bacterial growth inexpressed prostatic secretion (EPS) and post-prostate massage urine (VB3) samples but negative first-voided (VB1) and midstream urine (VB2) samples were included in the study. The Phoenix system (Becton Dickinson) was used for bacterial identification and antimicrobial susceptibility testing.Staphylococcus spp. were the most prevalent microorganisms in patients with chronic prostatitis (60.8%), followed by Escherichia coli and Enterococcus spp. (13.7%).Most patients with chronic prostatitis who were diagnosed with Staphylococcus spp. exhibited high resistance tobenzylpenicillin (75.0%), oxacillin (60.5%), and ampicillin (59.0%). Patients diagnosed with Enterococcus spp. showed high resistance to quinupristin-dalfopristin(20.0%), cefoxitin screen (15.7%), clindamycin (16.9%), tetracycline (16.9%), and rifampicin (17.1%). Among those diagnosed with Streptococcus spp., most had resistance to oxacillin (7.4%), tobramycin (8.0%), erythromycin (8.4%), clindamycin (8.4%), tetracycline (8.4%), and mupirocin (12.1%).The patients with E. coli had resistance to extended-spectrum β-lactamases (ESBL)(45.5%) and cefepime (25.0%). In summary, we found that the most prevalent pathogens from patients with chronic bacterial prostatitis are E. coli, Enterococcus spp., and Staphylococcus aureus. Gram-positive isolates showed the highest resistance to benzylpenicillin, fosfomycin, ampicillin, tetracycline, rifampicin, and mupirocin. Moreover, gram-negative bacteria were most resistant to ESBL, cefepime, and ampicillin-sulbactam. To manage this condition, physicians should take into consideration the development of multi-drug resistance among the pathogenic agents.