Objective: To investigate the positive rate of the clinical isolated enterococcus, distribution of samples and drug resistance transition, and to provide a guidance to clinical medication and prevention and control of the nosocomial infection. Methods: Clinical isolated enterococcus was collected from the monitoring data of antimicrobial resistance by the Affiliated Hospital of Xuzhou Medical University from 2014 to 2018. Identification of bacteria and the drug sensitive test were performed, the distribution of pathogenic bacteria and the drug resistance were analyzed, and the software of WHONET 5.6 and SPSS 15.0 were used to analyze the data. Results: A total of 1 214 lines of non-repetitive enterococcus were isolated from 2014 to 2018, and only enterococcus faecalis (573) and enterococcus faecium (641) were included, 16 of which were enterococcus resistant to vancomycin. The source of bacterial strain mainly came from urine. These two kinds of enterococcus remained a low level of resistance to vancomycin, teicoplanin, linezolid and tigecycline. Besides, enterococcus faecalis showed drug resistance rates of 80.6% and 63.1% for erythrocin and highly concentrated gentamicin, respectively, and the rates for other antibacterial were all below 53%, and enterococcus faecium showed a drug resistance rate of above 66% for other antibacterial. The drug resistance rate of enterococcus faecalis showed a decrease year by year from 1.6% to 0 and that of enterococcus faecium from 5.6% to 1.3%. Their difference showed no statistical significance, and P value were 0.350 and 0.245, respectively. Conclusion: The positive rate of enterococcus during the 5 years gradually increased from 1.7% in 2014 to 1.9% in 2018, with an uptrend, and the drug resistance to the antimicrobials showed certain difference, suggesting that the antimicrobials can be reasonably selected by the clinician to control the nosocomial infection.