Objective To investigate current situation of Chinese clinical microbiological laboratories participating in management of healthcare-associated infection and antimicrobial use. Methods Fourteen provinces (municipalities), autonomous regions and army hospitals in seven regions of China were selected, the participation of clinical microbiological laboratories in the consultation of HAI diseases, specimen quality control, antimicrobial use, and management of multidrug-resistant organisms (MDROs) before 2000 and every five years from 2000 to 2015 were investigated, the surveyed results were analyzed statistically. Results A total of 187 hospitals were investigated, in 2015, 96 and 172 hospitals (51.34%, 91.98%) participated in the consultation of infectious diseases and multi-department collaborative management on MDROs respectively. However, 44 hospitals (23.53%) still manually performed statistical analysis on drug susceptibility, only 26 hospitals (13.90%) had the ability of identifying homology of pathogens. Rate of MDRO surveillance data feedbacked to clinical departments increased from 66.84% (n=125) in 2010 to 95.72% (n=179) in 2015, the frequency of feedback was mainly monthly and quarterly; rate of antimicrobial susceptibility results feedbacked to clinic departments increased from 62.03% (n=116) to 94.12% (n=176), 82.35% (n=154) of clinical microbiological laboratories conducted quarterly feedback; the quality control rate of microscopic sputum smear before sputum culture increased from 63.10% (n=118) to 87.17% (n=163); rate of bilateral double blood culture increased from 35.83% (n=67) to 72.73% (n=136); rates of other aseptic body fluid culture (except blood and urine) increased from 4.86% to 5.74%; differences were all significantly different between 2010 and 2015 (all P<0.05). Conclusion Clinical microbiological laboratories have played an important role in promoting the development of HAI management in China, especially during the period of 2011-2015. However, the homology analysis on HAI pathogens, informatization of result feedback, and sterile body fluid specimens detection need to be further strengthened.