Objective To investigate changes in the distribution and antimicrobial resistance of bacteria isolated from cerebrospinal fluid (CSF) specimens in Hunan Province, and provide reference for correct clinical diagnosis and rational antimicrobial use. Methods Data reported by member units of Hunan Province Antimicrobial Resistance Surveillance System from 2012 to 2021 were collected according to China Antimicrobial Resistance Surveillance System (CARSS) technical scheme. Data of bacteria isolated from CSF specimens and antimicrobial susceptibility testing results were analyzed with WHONET 5.6 and SPSS 20.0 software. Results A total of 11 837 bacterial strains were isolated from CSF specimens from member units of Hunan Province Antimicrobial Resistance Surveillance System from 2012 to 2021. The top 5 strains were coagulase-negative Staphylococcus (n=6 397, 54.0％), Acinetobacter baumannii (n=764, 6.5%), Staphylococcus aureus (n=606, 5.1％), Enterococcus faecium (n=465, 3.9%), and Escherichia coli (n=447, 3.8%). The detection rates of methicillin-resistant coagulase-negative Staphyloco-ccus (MRCNS) and methicillin-resistant Staphylococcus aureus (MRSA) were 58.9%-66.3% and 34.4%-62.1%, respectively. No Staphylococcus spp. were found to be resistant to vancomycin, linezolid, and teicoplanin. The detection rate of Enterococcus faecium was higher than that of Enterococcus faecalis, and the resistance rates of Enterococcus faecium to penicillin, ampicillin, high concentration streptomycin and levofloxacin were all higher than those of Enterococcus faecalis (all P=0.001). Resistance rate of Streptococcus pneumoniae to penicillin was 85.0%, at a high level. Resistance rate of Escherichia coli to ceftriaxone was >60%, while resistance rates to enzyme inhibitors and carbapenem antibiotics were low. Resistance rate of Klebsiella pneumoniae to ceftriaxone was >60%, to enzyme inhibitors piperacillin/tazobactam and cefoperazone/sulbactam was >30%, to carbapenem imipenem and me-ropenem was about 30%. Resistance rates of Acinetobacter baumannii to most tested antimicrobial agents were ＞60%, to imipenem and meropenem were 59.0%-79.4%, to polymyxin B was low. Conclusion Among the bacteria isolated from CSF specimens, coagulase-negative Staphylococcus accounts for the largest proportion, and the overall resistance of pathogenic bacteria is relatively serious. Bacterial antimicrobial resistance surveillance is very important for the effective treatment of central nerve system infection.