[Objective] To evaluate the effect of eight aminoglycoside antibiotics against mycobacteria. [Methods] Minimal inhibitory concentration (MIC) of eight aminoglycoside antibiotics against twenty mycobacteria species in vitro were determined respectively. [Results] The various drugs show different antibacterial effect spectrum in vitro. [Conclusion] Aminoglycoside antibiotics are selection drugs in the treatment of nontuberculosis mycobacterial diseases clinically.