Abstract: OBJECTIVE To investigate the antimicrobial resistance and molecular epidemiological characteristics of K1 serotype Klebsiella pneumoniae strains from the hospitals in Huzhou. METHODS Totally 572 clinical isolates of non-repetitive K. pneumoniae were collected from 7 sentinel hospitals of Chinese Pathogen Identification Net in Huzhou. A total of 50 strains of K1 serotype K. pneumoniae were screened out by fluorescent quantitative polymerase chain reaction (PCR). The string test and antimicrobial susceptibility testing were performed. The molecular characteristics， including multilocus sequence typing (MLST)， drug resistance genes， virulence genes， plasmids and virulence scores were determined through whole genome sequencing analysis. The phylogenetic tree was constructed based on core gene single nucleotide polymorphism (cgSNP)， and the characteristics of the evolution of the serotype strains were observed. RESULTS The K1 serotype K. pneumoniae strains isolated from Huzhou accounted for 8.74%， the positive rate was 92.00% for the string test. The result of antimicrobial susceptibility testing showed that the drug resistance rate to ampicillin was highest (96.00%)， followed by chloramphenicol (56.00%) and nalidixic acid (42.00%). MLST analysis indicated that the 50 strains belonged to 6 ST types， and ST23 was the predominant type， accounting for 76.00%. The bioinformatics analysis showed that all of the strains carried 46 types of drug resistance genes in total， the carrying rate of β-lactamase gene bla_SHV-11 was highest (86.00%); the carrying rates of five types of virulence genes clbA， ybtS， iroB， iucA and rmpA were 84.00%， 96.00%， 100.00%， 94.00% and 96.00%， respectively. According to Kleborate database， totally 41 strains reached the highest virulence score. The phylogenetic tree showed that there were 7 evolutionary clusters. CONCLUSIONS The proportion of K1 serotype hypervirulent K. pneumoniae is high in Huzhou， ST23 is the predominant epidemic strain. The drug resistance is severe， and it is necessary to strengthen the surveillance and identify the potential risk of spread of infections.