Despite being classified as nosocomial pathogens, coagulase-negative staphylococci (CoNS) have long been considered as avirulent pathogens. CoNS are differentiated from the closely related but more virulent Staphylococcus aureus by their inability to produce free plasma coagulase. Currently, S. epidermidis is the most leading nosocomial pathogen among the CoNS, causing clinical implications and economic burdens. Among the essential virulence factors of staphylococci is beta hemolysin (hlb), an exotoxin responsible for cell disruption during bacteria invasions. The objective of the study was to isolate and clone beta hemolysin gene among isolates of S. epidermidis using species-specific primers and moreover, to test the hemolytic efficacy of the recombinant protein on blood agar. A total of seven S. epidermidis clinical isolates obtained from diabetic foot infection was used in this study. The hemolysin gene was amplified by polymerase chain reaction (PCR) and cloned into E. coli DH10B strain. The recombinant protein was then tested on sheep blood agar. The results showed the presence of beta hemolysin gene in all the seven S. epidermidis isolates tested. The efficacy of the recombinant gene to show hemolysis was found on the sheep erythrocytes. Clinical infections relating to CoNS have been increasing dramatically in the previous decades, and the virulence factors play essential role in the pathogenicity of these infections. Our results confirmed the importance of these cytotoxin genes in the development of infections caused by CoNS.