Prosthetic joint infections (PJIs) represent a severe complication in orthopaedics. Coagulase-negative staphylococci (CoNS) and Staphylococcus aureus represent the most frequent cause, but Gram-negatives have also been reported. With a view to describing the aetiology of PJIs diagnosed from January 2005 to September 2007 at S. Corona Hospital in Pietra Ligure, Italy, we conducted retrospective analysis of pathogens isolated from PJIs by means of surgical specimens, needle aspirates or swabs of fistula (3 samples). During the study period 228 PJIs were described and 141 (62%) were microbiologically documented and evaluated. Early and delayed infections represented 45% of episodes, while late infections were observed in 55%. The aetiology was mono-microbial in 84% of cases, and polymicrobial in 16%. CoNS and S. aureus were the most frequently isolated pathogens. In early and delayed infections methicillin resistant CoNS were 30% and 24%, respectively, while in late infections they were 17%. Methicillin-resistant S. aureus was isolated in 13% of early, 22% of delayed and 15% of late infections. Gram-negatives were described in 16% of episodes without differences being found in the three groups. In our report staphylococci represented the most frequent cause of PJIs. Methicillin-resistant strains were more frequently isolated in early and delayed infections, but their frequency in late episodes was not negligible. Polymicrobial infections and Gram-negative infections were also frequent.