Aims: Comparison of six commercially available in human medicine well-established slide agglutination systems for the identification of Staphylococcus aureus. Methods and Results: Slide agglutination tests were compared with the conventional tube coagulase test, biochemical identification and with the molecular identification by polymerase chain reaction (PCR) amplification of species-specific parts of the gene encoding the 23S RNA. Systems evaluated included Masta-Staph((R)) (Mast Diagnostics), Staphylase-Test((R)) (Oxoid), Staphytect-Plus((R)) (Oxoid), Staphyloslide Latex((R)) (Becton Dickinson), Slidex Staph Plus((R)) (bioMerieux) and Dry Spot Staphytect Plus((R)) (Oxoid). A total of 141 staphylococcal strains isolated from cases of bovine mastitis including 90 S. aureus, 14 Staphylococcus epidermidis, 10 Staphylococcus warneri, 13 Staphylococcus xylosus, 11 Staphylococcus haemolyticus and three other coagulase-negative staphylococci were tested with each method. Staphylococcus aureus strains were selected by macrorestriction analysis with pulsed field gel electrophoresis (PFGE). Only genetically unrelated strains were included in the study. The sensitivities and specificities of the test were as follows: Masta-Staph((R)) 86.7 and 90.1%, Staphylase-Test((R)) 78.4 and 85.1%, Staphytect-Plus((R)) 81.1 and 86.5%, Staphyloslide Latex((R)) 77.8 and 84.4%, Slidex Staph Plus((R)) 77.8 and 84.4%, Dry Spot Staphytect Plus((R)) 75.6 and 83.0%. Conclusions: The results of this evaluation suggest that the six slide agglutination methods tested can provide rapid identification of S. aureus also from bovine mastitis. The sensitivity and specificity seems to be less than those reported from human S. aureus isolates. Significance and Impact of the Study: This is one of the first comparative reported investigations about the applicability of different commercially available slide agglutination tests for the detection of S. aureus from bovine mastitis using PFGE selected clinical isolates.