The present study aims to identify the emergence of Methicillin-resistant Staphylococcus aureus, S. aureus resistant to vancomycin and to investigate the presence of Mec A, Van A and Van B genes among Staphylococcus strains isolated from hospital environment. For each type of sample, surface of beds, recyclable material, boxes of rodar, floor and door slats, 53 samples were taken. So, a total of 265 samples were collected by swabbing (except boxes of rodar) in a private clinic in southern Benin. Bacteriological analysis was performed using the conventional method followed by DNA extraction with the Quiagen kit. The resistance genes Mec A, Van A and Van B were sought using specific primers. 215 samples were culture positive with 155 strains (62%) of coagulase negative (CNS) staphylococci and 95 strains (38%) of S. aureus. The majority of strains were resistant to gentamycin and clindamycin. These 155 strains were carried the Mec A gene and 10 strains carried the Van A and Van B gene. The study reveals the presence of resistant Staphylococci carrying the Mec A gene, which could be responsible for nosocomial infections in patients. Hygiene must be improved to limit the spread of these germs and protect patients.