Objective To investigate the change of the break point of the CLSI in different years on the drug sensitivity of Enterobacteriaceae bacteria. This is valuable for rational use of antibiotics. Methods To retrospective analyze 2014 Enterobacteriaceae bacteria susceptibility variation by CLSI file cefepime breakpoint of 2013(S23) and 2014(S24). Results Compared with breakpoint of S23, E. coli and Klebsiella pneumonia, cefepime resistant rate rose by 2.6% and 2.8% respectively, and the resistance difference was statistically significant (P<0.05); ESBLs-producing E. coli, ESBLs-producing Klebsiella pneumoniae and ESBLs non-producing Klebsiella pneumoniae toword cefepime resistant rate rose by 3.8%, 2.4% and 2.0% respectively, and the resistance difference was statistically significant (P<0.05); For other Enterobacteriaceae bacteria, such as ESBLs non-producing E. coli and Enterobacter cloacae with cefepime in drug resistance rate had no significant difference (P>0.05). Compared with S23, variety in drug susceptibility of bacteria was most significant in S24, during cefepime MIC of 4 μg/mL, 8 μg/mL, 16 μg/mL. Conclusion According to the S24 breakpoint, the drug resistance rates of the bacteria in the intestines of the bacteria is increasing, especially in ESBLs strain; antibiotics use should be following laboratory results.