Abstract: OBJECTIVE To understand the current status of prevalence of multidrug-resistant organisms（MDROs） and use of antibiotics in second grade or above hospitals of Guizhou province so as to provide guidance for prevention and control of nosocomial infection. METHODS By means of questionnaire survey, the data of isolated MDROs and use of antibiotics were collected from the surveyed hospitals from Jan 1, 2017 to Dec 31, 2017, and the statistical analysis was performed with the use of SPSS 23.0 software. RESULTS The data of the 234 hospitals surveyed showed that the incidence of MDROs nosocomial infection was 0.076%, the isolation rate of MDROs was 14.04%; there was significant difference in the isolation rate among the MDROs（χ²=15478.703,P＜0.001）, the isolation rate of Carbapenem-resistant Acinetobacter baumanni（CRAB） was the highest（44.14%）, the isolation rate of Vancomycin-resistant Enterococcus faecalis was the lowest（2.39%）.There was significant difference in the incidence of nosocomial infection among the MDROs（χ²=1399.744,P＜0.001）,the incidence of CRAB nosocomial infection was the highest（0.0279%）, and the incidence of Vancomycin-resistant Enterococcus faecalis nosocomial infection was the lowest（0.00033%）.The utilization rate of antibiotics of the hospitalized patients was 50.90%, the etiological submission rate was 33.85% before the antibiotics were used.There were significant differences in the above indexes among the different grades of medical institutions（P＜0.001）. CONCLUSION The mastery of data of the second grade or above medical institutions of Guizhou provinces such as the isolation rate of MDROs and utilization rate of antibiotics may lay the foundation for prevention and control of nosocomial infection in the medical institutions under the instruction of quality control center.