Methicillin-resistantStaphylococcus aureus(MRSA) are spreading worldwide in hospital and community settings, thus posing a serious public health problem. Panton-Valentine Leukocidin (PVL), an important virulence factor ofS. aureus, is a marker of community-acquired MRSA. Here we determined the prevalence ofpvlgenes amongS. aureusisolates from different hospitals in the Gaza Strip, Palestine. A total of 285S. aureusisolates were collected from five different hospitals in the Gaza Strip. All isolates were characterized for their susceptibility patterns to available antimicrobial agents and by using multiplex PCR for the detection ofmecAandpvlgenes. The overall prevalence of MRSA in Gaza hospitals was 70.2% (range: 76.3% to 65.5%) and that ofpvlamongS. aureusisolates was 29.8% (range: 32.9% to 26.2%). Thepvlgene was equally prevalent among MRSA isolates (30.5%) and MSSA isolates (28.2%). The most effective antibiotics were rifampicin, vancomycin, and clindamycin, with susceptibility rates of 91.2%, 88.7%, and 84.6%, respectively. The highest percentage of strains were observed to be resistant to penicillin and amoxicillin with clavulanic acid—96.1% and 73.6%, respectively. Our results showed a high prevalence of MRSA andpvl-positive isolates in Gaza Strip hospitals, which likely reflects the situation in the community. It is mandatory to implement systematic surveillance of both hospital and community isolates, together with interventions (such as increased hand hygiene, use of hydroalcoholic solutions, and isolation of carriers) to limit their spread.