Objective To analyze the clinical characteristics and risk factors of extracorporeal membrane oxygenation (ECMO)-related healthcare-associated infection (HAI), provide theoretical basis for the prevention and control of ECMO-related HAI. Methods Patients who received ECMO treatment in a tertiary hospital from June 2014 to September 2018 were retrospectively analyzed, according to the 1∶2 matching method, patients who didn't receive ECMO treatment were selected as the control group (non-ECMO group), basic information and hospitalization expense of two groups of patients were compared. According to whether ECMO-related HAI occurred, patients who received ECMO treatment were divided into ECMO infection group and ECMO control group, clinical characteristics of two groups of patients were compared, risk factors, mortality and medical expenses of ECMO-related HAI were evaluated. Results A total of 30 patients who received ECMO treatment were included in the study, with an average age of (41.9±16.2) years, 76.7% of the patients were male, and 22 patients underwent venovenous ECMO (VV ECMO). 15 patients had 24 cases of ECMO-related HAI, including 12 cases of bloodstream infection, 9 cases of pulmonary infection, 1 case of urinary tract infection, 1 case of surgical site infection and 1 case of gastrointestinal infection. Patients in ECMO infection group had longer ECMO support time, higher proportion of tracheotomy and lower hemoglobin level. Multivariate logistic regression analysis showed that ECMO support time ≥ 10 days was an independent risk factor for ECMO-related HAI. Compared with ECMO control group, patients with ECMO-related HAI had longer hospital stay ([44.5±39.6] d vs [18.4±11.7] d, P=0.021) and higher medical expense ([413 359.9±204 887.0] Yuan vs [264 925.3±76 079.7] Yuan, P=0.004). Conclusion HAI is a common complication of ECMO treatment, which significantly increases medical expense and length of hospital stay. Defining indications of ECMO treatment as well as stopping respiratory circulatory support as early as possible can reduce the incidence of ECMO-related infection.