Abstract: In order to study the species and drug resistance of spore-forming bacteria,settled bacteria was selected from clean environment by the national standard(GB/T 16294-2010). The spore-forming strains were screened from the purified settled bacteria by the method of Schaeffer-Fulton’s spore dyeing. The 16S rDNA sequence alignment analysis was used to identify the strains. The method of K-B was used to detect the resistance of the strains to 20 antibiotics. The results showed that 74 strains were collected and 14 spore-forming strains were detected in different clean environments(pharmaceutical,clean bench,cell room),and the detection rate of spore-forming strains in clean environments was 18.9%. At the same time,it was found that its resistance rate to β-lactam antibiotics was 92.86%. It had shown that spore-forming strains in the clean environment had certain resistance and provided reference data for the safety risk of spore-forming strains in clean environment.