Abstract: OBJECTIVE To perform the high-throughput analysis of the bacterial pathogens causing infectious diarrhea in Huai'an City in 2024 so as to provide basis for development of targeted prevention and control strategies. METHDOS A total of 316 patients with infectious diarrhea who visited outpatient department of medical institutions in Huai'an City from May 2024 to Dec. 2024 were recruited as the research subjects. The bacterial pathogens causing the infectious diarrhea， epidemic features and molecular genetic characteristics of the main pathogens were analyzed by means of multipathogen nucleic acid screening， metagenomic next-generation sequencing (mNGS) and whole-genome sequencing analysis. RESULTS The total detection rate of nucleic acid of pathogens was 46.84%(148/316)among the 316 diarrhea samples， and the isolation rate of enteroaggregative Escherichia coli (EAEC) was highest (24.68%， 78/316). A total of 27 strains of EAEC， 11 strains of enteropathogenic E. coli (EPEC)， 7 strains of Salmonella， 5 strains of enterotoxigenic E. coli (ETEC) and 2 strains of Vibrio parahaemolyticus were isolated via culture. Among the 33 samples that tested negative by both nucleic acid screening and conventional culture， 4 were found to harbor pathogens associated with infectious diarrhea through metagenomic sequencing. The 2 V. parahaemolyticus isolates were identified as ST3 and ST1343， respectively; only 1 strain carried the tdh gene. The diarrheagenic E. coli isolates comprised 24 known ST types and 11 novel ST types， which did not show clustering on the whole-genome single nucleotide polymorphism (wgSNP) phylogenetic tree. The serotypes of Salmonella included S. enteritidis， S. typhimurium var. monophasic， and S. typhimurium， with close genetic relationship observed among the strains sharing the same serotype. CONCLUSIONS The spectrum of the bacterial pathogens causing infectious diarrhea is diverse in Huai'an City， with EAEC being the predominant pathogen. The combination of high-throughput screening technology and culturomics enables the highly efficient detection and traceability of the pathogens causing infectious diarrhea.