Chronic prostatitis (CP) or Persistent prostatitis affecting the male urinary system is the hallmark of Chronic Pelvic Pain Syndrome (CPPS), an illness that is sometimes challenging to diagnose and cure. Symptoms of CPPS include discomfort and lower urinary tract abnormalities. To determine the frequency of bacterial isolates causing prostatitis and their antimicrobial resistance profile, examine the spectrum and rates of resistance to antibacterial agents in the pathogens of bacterial prostatitis in patients, assess the occurrence of multidrug-resistant (MDR) cases, and identify different risk factors (age group) associated with prostatitis infection, the current study was designed. After clinical samples were cultivated, standard microbiological techniques were used to identify the bacterial isolates, which VITEK subsequently validated. 131 isolates from cultures of urine and seminal fluid from patients aged 25-85 years. The age range of 46–55 years old had the greatest number of positive cases. There were 26 (19.84%) Gram-positive and 105 (80.15%) Gram-negative pathogenic bacteria. While Staphylococcus aureus (73.07%) was the most common Gram-positive strain, Escherichia coli (E. coli) was the most isolated Gram-negative strain (41.9%). Gram-negative bacteria were highly resistant 100% to Cefuroxime, Amoxicillin+clavulanic acid and Amoxicillin due to their increased resistance to widely used antibiotics, these pathogens reduced the number of available treatments and increased the risk of recurring or chronic infections. This result emphasises the critical need for enhanced infection control methods, sensitivity-test-based customised antibiotic therapy, and ongoing antimicrobial surveillance. However, co-trimoxazole was not effective against Gram-positive bacteria. Imipenem, Erythromycin, Pristinamycin, Chloramphenicol, Rifampicin and Ciprofloxacine increase the risk of persistent or recurrent infections and emergence of multidrug-resistant Staphylococcus aureus and Enterococcus spp. Targeted interventions are critical to manage resistance and improve patient outcomes in chronic prostatic infections. In order to control resistance and enhance patient outcomes in cases of chronic prostatic infections, targeted therapies are essential.