ObjectiveTo investigate the causes of clustered event of acute hemorrhagic conjunctivitis(AHC) in the ophthalmology department of a hospital, and provide basis for comprehensive prevention and control of healthcareassociated infection(HAI). MethodsAHC occurred in the ophthalmology department of a hospital from January to November 2016 were investigated retrospectively, onsite epidemiological and environmental hygienic investigation was carried out. Hand hygiene compliance among health care workers(HCWs) in this department during JanuaryAugust 2016(before supervision)  and SeptemberNovember 2016 (after supervision) were surveyed. Hands of HCWs, instruments for routine diagnosis, and frequently touched object surface were taken sampling by ATP fluorescence rapid detector, bundle intervention strategies for HAI prevention and control were formulated. ResultsA total of 11 patients developed AHC from August 4, 2016 to September 17, 2016, 9 of whom were inpatients, and 2 were outpatients; 3 HCWs had AHC. Eye secretion was taken only from two patients in ward I, virological examination confirmed that it was caused by adenovirus infection. ATP examination showed that the qualified rate of HCWs’ hand hygiene and hygiene of environmental object surface were 28.57% and 34.38% respectively. Compliance rates of hand hygiene of HCWs before and after supervision were 50.00% and 78.52% respectively, difference was statistically significant (χ2=4.89, P=0.03). During the supervision period, the implementation rate of disinfection for equipment such as slit lamp, tonometer, and other common equipment was 76.64%. After the implementation of bundle intervention strategies, no case of hemorrhagic conjunctivitis was found in the ophthalmology department since midOctober. ConclusionGood hand hygiene and environmental cleaning and disinfection are still important means of infection prevention and control, timely and feasible bundle intervention strategies can effectively control the outbreak or epidemic of infection.