The application of effective disinfectants during experimental activities of high­pathogenic microorganism is crucial for decontamination of laboratory environment to prevent laboratory­acquired infections. In this study, the quantitative germicidal disinfectant validation experiments using membrane filtration suspension were conducted against Mycobacterium tuberculosis (M. tuberculosis) H37Rv. Sodium hypochlorite solution (containing 1 000 mg/L active chlorine), 75% ethanol/water, and Safespore disinfectant killed M. tuberculosis efficiently. When the concentration of ethanol decreased to 37.5%, it was still able to kill M. tuberculosis by prolonging killing time to 10 min. The results suggest that 75% ethanol/water is suitable as conventional disinfectant and sodium hypochlorite solution (containing 1 000 mg/L active chlorine) is suitable under emergency and accident in biosafety level 3 laboratories (BSL­3)．