〔Abstract〕Objective: To analyze the distribution characteristics and drug resistance of the blaSHV, blaTEM, blaCTX-M-3, and blaCTX-M-14 genes in extended-spectrum β-lactamase Enterobacteriaceae (ESBL-E), and to provide a basis for the rational use of antibacterial drugs and infection prevention and control. Methods: From August 2023 to April 2024, 332 Enterobacterales isolates were collected from a hospital in Dali Bai Autonomous Prefecture. ESBLs⁃positive isolates were identified by phenotypic confirmatory tests following CLSI guidelines. Genotypes were analyzed using PCR amplification, agarose gel electrophoresis and gene sequencing, and antimicrobial resistance profiles were assessed by susceptibility testing. Results: The detection rate of ESBL-E was 30.1% (100/332). Among these, Escherichia coli had the highest ESBLs positivity rate (74/208, 35.6%), followed by Klebsiella pneumoniae (20/90,22.2%). The detection rates of blaCTX-M-14 , blaCTX-M-3 , blaTEM , and blaSHV genes were 54%, 45%, 43% and 21%, respec⁃ tively. Single genotypes accounted for 50%, predominantly blaCTX-M-14 and blaCTX-M-3 . Among mixed genotypes, blaSHV and bla⁃ TEM were all co-expressed with blaCTX-M (dual genotypes 30%, triple genotypes 19%). Among isolates carrying blaCTX-M type genes, blaCTX-M-14 was predominant in purulent secretions (76 .2%, 32/42); in urine, blaCTX-M-3 (59 .5%, 22/37) and blaCTX-M-14 (48 .6%, 18/37) showed co-prevalence; in lower respiratory tract specimens, blaCTX-M-3 was overwhelmingly dominant (93 .3%, 14/15). ESBL-E exhibited resistance rates > 70% to ampicillin, piperacillin, cefazolin, cefotaxime, cefepime, quinolones (ciprofloxacin, le⁃ vofloxacin), tetracycline, and trimethoprim/sulfamethoxazole. Conclusion: In a hospital of Dali Bai Autonomous Prefecture, ESBL-E are predominantly E. coli carrying the single genotype blaCTX-M-14 or blaCTX-M-3 . The blaSHV and blaTEM genes are often coexpressed with blaCTX-M. Empirical therapy should consider the infection site and local genotype prevalence , indicating the need to strengthen genotype surveillance to guide precise antimicrobial use.