Objective:To understand the rotavirus contamination in the environment of microbiology laboratory, identify the key parts prone to contamination in the laboratory, and evaluate the inactivation efficacy of three common disinfectants on rotavirus.Methods:The laboratory environment and the surface of personal protective equipment were sampled with virus sampling swab after the experimental operation, and rotavirus nucleic acid was detected by real-time RT-PCR. Rotavirus inactivation experiments were carried out on chlorine containing disinfectant, ethanol and benzalkonium chloride according to the Chinese Technical Standard of Disinfection (2002 version).Results:A total of 174 samples were collected and 13 rotavirus nucleic acid positive samples were detected. The overall positive rate was 7.47%. The rotavirus could be inactivated with more than 4.00 log titer reduction, with 500 mg/L sodium hypochlorite disinfectant for 5 minutes, 75% ethanol for 1 minute and 1 500 mg/L benzalkonium chloride for 1 minute.Conclusions:There is a risk of rotavirus contamination in the microbiology laboratory. Sodium hypochlorite, ethanol and benzalkonium chloride disinfectants can effectively inactivate rotavirus and can be used for rotavirus decontamination.