Objective To investigate and control a suspected outbreak of Enterobacter cloacae(E. cloacae) bloodstream infection in hemodialysis patients in department of nephrology of a tertiary hospital, provide reference for clinical prevention and treatment. Methods Epidemiological investigation was carried out on hemodialysis patients in department of nephrology of a hospital from May 21 to June 6, 2018. Environmental hygiene detection were performed on hospital air, dialysis water and dialysate, dialysis materials, in-use disinfectant, hands of health care workers(HCWs), object surface, as well as hands and skin at deep vein catheterization sites of patients, causes of infection were analyzed, strict prevention and control measures were taken. Results From May 21 to June 6, 2018, a total of 6 patients suffered from catheter-related bloodstream infection(CRBSI), all were isolated E. cloacae from blood culture, incidence of CRBSI was 3.03‰(6/1 980). 133 spcimens were collected for environmental hygiene monitoring, detection results of air, dialysis water and dialysate, disposable dialysis pipeline, and in-use disinfectant were all qualified; qualified rate of hand sampling of HCWs was 61.54% (8/13); qualified rate of object surface sampling was 80.28% (57/71), 6 specimens of hands and skin at deep venous catheterization sites of patients were taken, 1 specimen was unqualified; E. cloacae was not detected in environmental hygiene monitoring. Risk factors analysis showed that incidence of E. cloacae CRBSI was higher in patients with diabetes mellitus and those who needed thrombolysis before dialysis, difference were both statistically significant (both P<0.05). After 2-week treatment during hospitalization, blood cultures of 6 infected patients were all negative, patients continued hemodialysis treatment in outpatient departmen after discharged. No new infection case was observed in the blood purification center for 2 months. Conclusion Blood purification center should strengthen hand hygiene of HCWs, clean and disinfect environment, and cut off the route of exogenous infection. Strengthening the management of vascular catheterization site and blood sugar of hemodialysis patients, as well as avoidance of thrombolysis before hemodialysis can reduce the occurrence of CRBSI.