In order to investigate the clinical disinfection effect of glutaraldehyde-decamethyl ammonium bromide atomizing on porcine reproductive and respiratory syndrome virus（PRRSV），Streptococcus suis and porcine Escherichia coli（E. coli），based on laboratory conditions，the disinfection method was simulated to spray glutaraldehyde decamethyl ammonium bromide and suspension synergist into a disinfected space in the form of mist，and to disinfect the culture dish coated with PRRSV and the serum medium inoculated with Streptococcus suis and E. coli. After disinfection，the virus was eluted from the culture dish and inoculated into Marc-145 cells，then the serum medium was placed in an appropriate environment for incubation，and cell pathological changes and bacterial growth were observed to identify the disinfection effect. Results showed that PRRSV could be disinfected to some extent when glutaraldehyde-decamethyl ammonium bromide was at a spatial concentration of 2.1 mL/m3 for 30 min；and completely inactivated at a spatial concentration of 3.5 mL/m3 for 60 min；Streptococcus suis could be completely inactivated at a spatial concentration of 2.1 mL/m3 for 50 min，and E. coli for 35 min. In conclusion，PRRSV，Streptococcus suis and E. coli could be effectively disinfected by the glutaraldehyde-decamethyl ammonium bromide atomizing that could be used for daily disinfection of empty pig houses in farms，indicating an important significance to the biological safety concerning disease prevention and control in the farms.