Choosing the appropriate method for differentiation of Staphylococcus aureus strains is important for effective diagnostics and epidemiological investigations. Despite the fact that the results of phenotypic methods are strongly dependent on environmental conditions, they can still be useful in the investigation of epidemic strains of S. aureus. In this article, the potential application of commonly used phenotypic methods in epidemiological studies of S. aureus was analysed. Advantages and disadvantages of methods such as biotyping, serotyping, phage typing, AST (Antimicrobial Susceptibility Testing), SDS-PAGE (Sodium Dodecyl Sulphate Polyacryl Gel Electrophoresis), MLEE (Multilocus Enzyme Electrophoresis) and MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectroscopy) were also discussed. Finally, phenotypic techniques were compared in terms of their discriminatory potential, typeability of isolates, time of analysis, reproducibility, ease of performance and ease of results interpretation.