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In order to provide useful information for effective control and clinical therapy of infection, the resistance status and the rate of carrying AmpC β-1actamase of Enterobacter cloacae (E. cloacae) were investigated. By VITEK (Bacterial automatic biochemical analyzer), the isolates of E. cloacae were identified and the drag resistance was measured. The AmpC enzyme was detected by the five-disk diffusion test. Antibiotic sensitivity test showed that the resistance effects of E. cloacae to cefazolin, cefoxitin and ampicillin were more serious, with resistant rates of 80.5%, 75.3% and 70.1%, respectively. However, it was more sensitive to Sulperazone (cefoperazone/sulbactam, 13.0%), amikacin (16.9%) and ciprofloxacin (19.5%). Meanwhile, the phenotype detection showed that 35.06% (27/77) isolates of E. cloacae produced AmpC β-1actamase. Most of E. cloacae are multi-drug resistant strains. Sulperazone (cefoperazone/sulbactam), a kind of component β-1actamase, is a more effective antibiotic for treating infection caused by E. cloacae. Unreasonable application of the third generation cephalosporins plays an important role in leading to emergence of high-yield AmpC β-1actamase strains, so antibiotics should be used wisely.