The virulence of five different concentrations of isolate of Beauveryia bassiana named MZ041016 against adult, larvae and pupae of Bactrocera dorsalis(Hendal) was tested in laboratory. Large amount of Bactrocera dorsalis(Hendal) of each stage were killed at the concentration 3.6×105 spores/mL.The mortality of 3.6×105 spores/mL against adult was 95.45%, and the lethal time(LT50) of this concentration was 3.562 d.The lethal concentration(LC50) of eight day after treatment was 2.14×105 spores/mL. The mortality of 3.6×108 spores/mL against larvae was 95.94%, and the lethal time(LT50) of this concentration was 4.926 d.The lethal concentration(LC50) of eight day after treatment was 3.03×105 spores/mL. The mortality of 3.6×108 spores/mL against pupae was 78.15%, and the lethal time(LT50) of this concentration was 5.358 d.The lethal concentration(LC50) of eight day after treatment was 3.31×105 spores/mL. The isolate of Beauveryia bassiana named MZ041016 had virulence against adult, larvae and pupae Bactrocera dorsalis (Hendal) in laboratory, and the application prospect of it in field should be feasible. The result provided a scientific basis for the pollution-free production of fruit and biology control of Bactrocera dorsalis (Hendal) in field.