Abstract: Objective The present investigation was conducted to identify the serological and biochemical characteristics of an non-Shiga toxin-producing Enterohemorrhagic Escherichia coli O157:H7 and its gene of toxicity. Methods The strain 154 was identified with routine serological and biochemical assays,and PCR,southern blot and Hela cell cytotoxicity assay were conducted to identify the specific gene of strain 154. Results PCR,southern blot and Hela cell cytotoxicity assay for Shiga toxin were negative,toxic factors of eae and hly were tested positive. Conclusion Strain 154 is an non-Shiga toxin producing Enterohemorrhagic Escherichia coli O157:H7 strain with specific toxic factors of O157:H7.