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Staphylococcus aureus (S. aureus), a vital nosocomial pathogen, is responsible for several diseases. With the increasing isolation rate in clinical specimens, rapid identification of this bacterial species is required. But present identification via conventional methods is time-consuming and lacks accuracy. The purpose of the current study was to evaluate the use of surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF MS) for rapid identification of S. aureus.