Abstract: OBJECTIVE To investigate the extended-spectrum β-lactamases(ESBLs)-producing Enterobacteriaceae isolated from the intestinal tracts of the preschool children in Wuxi and analyze the characteristics of drug resistance. METHODS Totally 237 stool samples were collected from the preschool children of two kindergartens in Wuxi City from Sep. 2024 to Nov. 2024. The specimens were cultured for isolation by ESBLs chromogenic media, the drug resistance was analyzed by drug susceptibility testing, and the carrying rates of virulence genes and molecular characteristics were analyzed by means of whole genome sequencing. RESULTS The isolation rate of ESBLs-producing Enterobacteriaceae was 59.92%(142/237) among the preschool children in Wuxi City; totally 157 strains involving 7 species, 5 genera were identified, the Escherichia(84.71%,133/157) was dominant, followed by Klebsiella(9.55%,15/157). The drug resistance rates of the strains to ampicillin, cefotaxime and tetracycline were 100.00%(157/157), 93.63%(147/157) and 70.06%(110/157), respectively; the multidrug-resistant rate reached up to 94.90%(149/157). The genomic sequencing analysis showed that the strains carried 96 types of drug resistance genes in 12 categories; the carrying rates of tet(A)(56.05%,88/157),mph(A)(50.32%,79/157) and sul1(43.95%,69/157) were relatively high. Totally 5 strains of Escherichia coli were resistant to carbapenems, 2 strains were resistance to polymyxin, and the strains carried high-risk drug resistance genes such as bla_NDM and mcr-1. ST types were diverse, ST38 and ST131 were more common. The strains generally carried multiple virulence genes, and the detection rates of entB and ompA were highest(99.36%, 156/157). CONCLUSIONS The carrying rate of ESBLs-producing strains is high in the intestinal tracts of health children in Wuxi City. Some of the strains are resistant to the key antibiotics and carry the high-risk drug resistance genes, indicating that the population of children may become a major reservoir for drug-resistant bacteria. It is necessary to strengthen the surveillance of drug-resistant strains and management of antibiotics.