Objective To investigate and analyze an outbreak event of suspected rotavirus healthcare-associated infection (HAI), provide reference for the prevention and control of HAI. Methods Using on-site epidemiological investigation method, 6 children with rotavirus infection in a tertiary hospital from March 26 to April 5, 2021 were investigated, environmental hygiene was monitored, the main relevant factors were analyzed, and effective control measures were taken. Results From March 26 to April 5, 2021, 59 patients were admitted to department of pedia-tric internal medicine for more than 48 hours, 6 patients were infected with rotavirus, incidence was 10.17%. Among 6 children, 4 were males and 2 were females; the mean age was (1.61±1.21) years; admission time of children was on March 19-30, all children were isolated rotavirus on the 5th to 7th day of admission, 6 children were in the same nursing group, beds 22, 23 and 25 were in the same ward. Rotavirus screening was performed on the surface of environmental objects, hands of health care workers(HCWs) and caregivers of 6 children during hospita-lization period, a total of 22 specimens were taken, 20 were with negative and 2 were with weakly positive (bedside tables and bed rails) results, isolation rate was 9.09%. The event lasted for 11 days, and no relevant cases were found since the control measures were taken on April 5. Conclusion This event is a suspected outbreak of HAI caused by rotavirus infection in the department of internal medicine, infection is timely and effectively controlled through strengthening the management of patients and caregivers, stabling medical treatment personnel, strengthening the cleaning and disinfection of ward environment, improving the compliance and correctness of hand hygiene of HCWs, and paying attention to cleaning process.