Abstract: OBJECTIVE To make clear of the clinical distribution and drug resistance spectrums of linezolid-non-sensitive Enterococci (LNSE) strains. METHODS The non-repetitive clinical isolates of LNSE were retrospectively collected from Beijing Chao-Yang Hospital, Capital Medical University between Jan. 2013 and Dec. 2022. The sources, department distribution and drug susceptibilities of the strains were observed. RESULTS A total of 103 strains of LNSE were isolated, there were 83 strains of Enterococcus faecalis, 16 strains of Enterococcus faecium, 2 strains of Enterococcus gallinarum, and 2 strains of Enterococcus casselifavus. The isolation rate of linezolid-non-sensitive E. faecalis was 3.77%, higher than 0.88% of the linezolid-non-sensitive E. faecium; the isolation rate of LNSE strains was higher after 2020 than before 2020 (χ²=38.230,P＜0.001). Among all the strains isolated from the departments, 35.35% were isolated from outpatient department, 17.17% were isolated from urology department, and 16.16% were isolated from critical care medicine department. The LNSE strains were mainly isolated from the hospitalized patients and outpatient department patients of the urology department. 60.61% of the strains were isolated from the male patients, and the patients aged more than 60 years old were dominant, accounting for 46.46%. The result of antimicrobial susceptibility testing showed that the drug resistance rates of the linezolid-non-sensitive E. faecalis strains to tetracycline, erythromycin, ciprofloxacin and levofloxacin were more than 50%, the drug resistance rates of the linezolid-non-sensitive E. faecium strains to penicillin, ampicillin, ciprofloxacin and levofloxacin were more than 90%, and 2 strains were detected to resistant to vancomycin. CONCLUSIONS The isolation rate of LNSE strains shows an upward trend in the hospital from 2013 to 2022, and the isolation rate is remarkably high after 2020. The E. faecalis strains are dominant and mainly isolated from urine specimens, and the urology department is the key department for prevention and control.