Screening for colonization with methicillin resistant Staphylococcus aureas (MRSA) is a key aspect of infection control to limit the nosocomial spread of this organism. Current methods for the detection of MRSA in clinical microbiology laboratories using conventional methods is time consuming. In this research we are trying to evaluate the use of real time PCR for the detection of MRSA. The PCR assay was evaluated in clinical isolates of MRSA (n = 45) and methicillin susceptible Staphylococcus aureas MSSA (n = 10). The diagnostic values of the assay showed high sensitivity and specificity. This real-time PCR assay proved to be a fast, sensitive and specific tool for MRSA detection in a routine microbiological laboratory. Real-time PCR now is available in all laboratories so its use in identification of MRSA will help in shortening the period for MRSA identification and will help in the success of infection control programs in hospitals.