The in vitro activity of levofloxacin against coagulase-negative staphylococci (CNS) was investigated. In vitro, on the basis of MIC90 values, levofloxacin was as active as ciprofloxacin, inhibiting both methicillin-sensitive and -resistant staphylococci at 0.5 μg/ml. The frequency of one-step development of levoflox-acin-resistant CNS was < 1 x 10––9. After repeated transfer of CNS in the presence of increasing concentrations of levofloxacin or other quinolones, there were 4-fold increases in MIC50 values for both levofloxacin and ofloxacin, whereas there were 64- to 128-fold increases for ciprofloxacin, norfloxacin and enoxacin. Based on MIC50 values, the concentrations of 1 or 2 ug/ml are clinically relevant. The mutant strains induced by levofloxacin showed less virulence based on the LD50 value in acute systemic murine infection.