Abstract: OBJECTIVE To investigate the clinical characteristic, drug resistance, virulence and molecular epidemiological characteristics of Klebsiella pneumoniae isolated from cerebrospinal fluid. METHODS A total of 21 K. pneumoniae strains isolated from cerebrospinal fluid specimens at the First Affiliated Hospital of Nanchang University from Aug. 2021 to Aug. 2024 were collected. Clinical data of patients with infections, drug susceptibility test results and polymerase chain reaction(PCR) amplification for drug resistance and virulence genes were retrospectively analyzed. Multilocus sequence typing(MLST), capsular serotyping(K typing) and pulsed-field gel electrophoresis(PFGE) were used to analyze their homology. RESULTS The 21 K. pneumoniae isolates from cerebrospinal fluid were from the neurosurgery department and ICU. Most infected patients had underlying intracranial diseases and underwent cranial surgery, with an overall poor prognosis rate of 76.19%. Laboratory tests showed elevated peripheral blood inflammatory markers, cerebrospinal fluid white blood cells and protein levels. K. pneumoniae strains were sensitive to ceftazidime-avibactam and showed relatively high sensitivity to tigecycline and colistin, while the resistance rates to β-lactams, other β-lactamase/β-lactamase inhibitors and carbapenem antibacterial agents were all 76.19%. Among the 21 K. pneumoniae strains, 16 carbapenem-resistant K. pneumoniae(CRKP) strains carried the KPC-2 carbapenemase resistance gene. And 12 carbapenem-resistant hypervirulent K. pneumoniae(CR-hvKP) strains were identified as ST11-K64/25, 4 CRKP strains were ST15-K19, and 3 hvKP strains(1 ST1265-K1, 1 ST23-K1 and 1 ST29-K54) were detected. CONCLUSIONS Patients with K. pneumoniae isolated from cerebrospinal fluid have various intracranial diseases and invasive procedures, accompanied by multiple complications and a poor prognosis. The predominant strain is ST11-K64/K25 CR-hvKP, suggesting that clinical monitoring and prevention of this high-risk clone should be strengthened.