| Summary: | Methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) is a common cause of severe surgical site infections (SSI). The molecular epidemiology of MRSA is poorly documented in Ethiopia. This study is designed to determine the prevalence of MRSA and associated factors among patients diagnosed with SSI. A multicenter study was conducted at four hospitals in Ethiopia. A wound culture was performed among 752 SSI patients. This study isolated <i>S. aureus</i> and identified MRSA using standard bacteriology, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS), and cefoxitin disk diffusion test. The genes <i>mecA</i>, <i>femA</i>, <i>vanA</i>, and <i>vanB</i> were detected through PCR tests. <i>S. aureus</i> was identified in 21.6% of participants, with 24.5% of these being methicillin-resistant Staphylococci and 0.6% showing vancomycin resistance. Using MALDI-TOF MS for the 40 methicillin-resistant Staphylococci, we confirmed that 31 (77.5%) were <i>S. aureus</i>, 6 (15%) were <i>Mammaliicoccus sciuri</i>, and the other 3 (2.5%) were <i>Staphylococcus warneri</i>, <i>Staphylococcus epidermidis</i>, and <i>Staphylococcus haemolyticus</i>. The gene <i>mecA</i> was detected from 27.5% (11/40) of Staphylococci through PCR. Only 36.4% (4/11) were detected in <i>S. aureus</i>, and no <i>vanA</i> or <i>vanB</i> genes were identified. Out of 11 <i>mecA</i>-gene-positive Staphylococci, 8 (72.7%) were detected in Debre Tabor Comprehensive Specialized Hospital. Methicillin-resistant staphylococcal infections were associated with the following risk factors: age ≥ 61 years, prolonged duration of hospital stay, and history of previous antibiotic use, <i>p</i>-values < 0.05. Hospitals should strengthen infection prevention and control strategies and start antimicrobial stewardship programs.
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