OBJECTIVE: To establish production management and the methods of microbiological quality control on breeding SPF chickens raised in the isolator. METHODS: The sera obtained from 10, 20, 40-week-old SPF chickens including introduced and self-breeding ones were used to monitor microbiological quality in order to detect the effectiveness of the current production management and the methods of microbiological quality control on SPF chickens. RESULTS: Because the hatching process was not strickly controlled during the early stage of breeding SPF chickens in the isolator, there were the emergence of microbiological contamination. Microbiological condition of the isolator breeding SPF chickens was effectively controlled after improving the operation management during the hatching process. CONCLUSION: If the disinfection during all breeding stages is strengthened, the microbiological quality of SPF chickens in the isolator could be ensured. The appropriate control on the density of SPF chickens in the isolator is not only beneficial to the breeding of SPF chickens and the recycling of hatching eggs, but also helpful to the control of environmental microbiology.