Aminoglycosides still play an important role in antistaphylococcal therapies, although emerging resistance amongst staphylococci is widespread. To further our understanding of the prevalence of aminoglycoside resistance in Europe, we tested 699 and 249 consecutive unrelated clinical isolates of Staphylococcus aureus and coagulase-negative staphylococci (CNS), respectively, from the SENTRY Antimicrobial Surveillance Program, for susceptibility to gentamicin, tobramycin, kanamycin and streptomycin, and examined the relationship between susceptibility to these antimicrobials and susceptibility to methicillin. Three hundred and sixty-three staphylococcal isolates demonstrated resistance to at least one of the aminoglycosides tested; all of these isolates were screened for the presence of aac(6')-Ie + aph(2"), ant(4')-Ia and aph(3')-IIIa, the genes encoding the most clinically relevant aminoglycoside-modifying enzymes. S. aureus isolates derived from hospital-acquired pneumonia tended to be more resistant to aminoglycosides and methicillin than isolates from blood or wound infections. In S. aureus, resistance to aminoglycosides was closely associated with methicillin resistance. Susceptibility of S. aureus to gentamicin has decreased by 9% from previous European studies to a current level of 77%, while susceptibility of CNS, currently at 67%, has increased by 21%. Geographical variation occurred, correlating with methicillin resistance, although intra-country variation was considerable. aac(6')-Ie + aph(2"), ant(4')-Ia and aph(3')-IIIa were found throughout Europe in 68%, 48% and 14% respectively of staphylococci resistant to at least one aminoglycoside. aph(3')-IIIa was considerably more common in methicillin-susceptible S. aureus and CNS isolates; the reverse was true for the other two resistance genes. The prevalence of ant(4')-Ia and aph(3')-IIIa genes in aminoglycoside-resistant staphylococci was significantly greater than that reported in previous European studies.