The increasing resistance of staphylococci to ß -lactam antibiotics has become a major clinical problem. Development of rapid and sensitive techniques for detection of MRSA is an important aim for public health. A duplex PCR were established for specific identification of methicillin-resistance Staphylococcus aureus (MRSA) in clinical samples. In this work a duplex SYBR Green real time PCR was developed for rapid identification of MRSA in local methicillin-resistance S. aureus isolates. Twenty methicillin-resistance S. aureus isolates, as determined by disc diffusion method, were subjected to DNA extraction and PCR amplification. Two genes were amplified successfully, mecA (533bp) and femA (314bp), as targets for methicillin-resistance and specific identification of S. aureus, respectively using conventional PCR. Sensitivity of the duplex PCR showed that the minimum concentration of DNA that gave positive results for the two genes was 30ng/µl. In order to develop rapid and sensitive test for identification of MRSA, serial dilutions of purified DNA were amplified gradually according to their concentrations using SYBR Green real time PCR. These results indicated that the SYBR Green real time PCR can be used for identification of methicillin-resistance S. aureus (MRSA) in clinical

Staphylococcusaureusis avirulence pathogenic bacterium.Detection of methicillin-resistant S. aureus(MRSA) using conventional culture and biochemical methods is labor and time consuming. Many MRSA isolates are heterogeneously resistant to ß-lactams, for example only 1 daughter cell out of 10 4 to 10 6 cells appears phenotypically resistant when routine antimicrobial susceptibility tests are performed. In this work two genes were used for detection of MRSA in urine, boil and nasal swabs using conventional PCR, mecA gene (533bp) were used for detection of methicillin resistant and femA gene (318bp) were used for S.aureus identification. It was found that not all resistant S. aureus tested with disk diffusion method carry the mecA gene that caused the resistant phenomena there were only 55% of all MRSA carrying mecA gene while femA gene gave 100% positive for S. aureus and this will lead us to appoint that penicillin-bindingprotein 2a (PBP2a) produced by mecA gene is not the only cause of resistant phenomena.