Methicillin-resistant Staphylococcus aureus (MRSA) is widely recognized as a significant pathogen responsible for numerous community-acquired infections across the globe. This study aimed to investigate the prevalence of Staphylococcus aureus, including MRSA strains, as well as their antimicrobial susceptibility patterns among pregnant women attending antenatal clinics at Mile Four Hospital and Maternity in Ebonyi State. A total of 150 urine specimens were collected from the participants and delivered to the Microbiology Laboratory of Ebonyi State University within two hours for laboratory examination. Upon arrival, the samples were initially inoculated into nutrient broth and incubated overnight to encourage bacterial growth. The enriched cultures were then subcultured on mannitol salt agar (MSA) plates and incubated aerobically at 37 °C for 24 hours to facilitate the isolation of S. aureus. The bacterial isolates were enumerated in accordance with the guidelines provided by the Clinical and Laboratory Standards Institute (CLSI), and their antimicrobial susceptibility was determined using the standard disc diffusion method. Of the 150 urine samples analyzed, 61 (40.7%) yielded Staphylococcus aureus. Among these isolates, 22 (36.0%) were identified as methicillin-resistant strains. The occurrence of both S. aureus and MRSA infections was most prominent among pregnant women aged 33–37 years and 38–42 years, with prevalence rates of 66% and 100%, respectively. Among occupational groups, civil servants showed the highest S. aureus prevalence (53%), while self-employed women had the highest MRSA prevalence (37%). Educational level also influenced prevalence, with S. aureus highest in women with secondary education (51%) and MRSA highest in those with primary education (45%). Third-trimester pregnant women showed a prevalence of 57% for both S. aureus and MRSA. The antimicrobial susceptibility analysis indicated that the Staphylococcus aureus isolates demonstrated substantial resistance to the majority of the antibiotics evaluated. High resistance rates were observed against oxacillin (97%), ceftriaxone (100%), gentamicin (97%), clindamycin (100%), tetracycline (92%), cefotaxime (100%), erythromycin (100%), vancomycin (100%), and ciprofloxacin (95%). In contrast, imipenem exhibited comparatively better effectiveness, with approximately 61% of the isolates showing susceptibility to this antibiotic. These findings are concerning, as most of these resistant antibiotics are widely used in hospitals and available on the market, highlighting an urgent need for monitoring and effective antibiotic stewardship
