We have shown previously that pls, which codes for the surface protein Pls of methicillin-resistant Staphylococcus aureus (MRSA), reduces adhesion to immobilized fibronectin, fibrinogen, laminin, and immunoglobulin G as well as invasion of host cells. Here, we tested a collection of 66 clinical MRSA isolates--48 negative for pls/Pls (pls(-)/Pls(-)), 15 positive for pls/Pls (pls(+)/Pls(+)), and 3 harboring the pls gene but not expressing Pls (pls(+)/Pls(-))--for cellular invasiveness. Invasion of 293 cells by pls(+)/Pls(+) strains was lower than that by the pls(-)/Pls(-) strains (median [range], 36% [22%-70%] vs. 93% [25%-162%]). The 3 pls(+)/Pls(-) strains (median [range], 95% [63%-103%]) were as invasive as the pls(-)/Pls(-) strains. In addition, we identified a pls(+)/Pls(+) staphylococcal chromosomal cassette mec (SCCmec) IV strain. In conclusion, 3 properties--pls/Pls, SCCmec type, and spa type--strongly predicted the cellular invasiveness of MRSA strains, as indicated by good clustering. In contrast, the spa type-deduced multilocus sequence typing clonal complex (MLST-CC) was not able to predict the invasiveness of MRSA strains equally well. The underlying mechanism remains to be elucidated.