Abstract:ObjectiveTo study the resistance to commonly used antimicrobial agents and ampicillin resistance mechanism of Haemophilus influenzae (H. influenzae). MethodsAntimicrobial resistance and βlactamase of H. influenzae isolated from inpatients and outpatients in a hospital from January 1 to December 31, 2012 were detected by BauerKirby disc diffusion method and nitrocefinbased test, TEM and ROB βlactamase genes were amplified by polymerase chain reaction (PCR). ResultsOf 100 patients infected with H. influenzae, 61.00% were aged 1-10 years. The resistance rate to ampicillin was 35.00%(n=35),resistance to sulphamethoxazole/trimethoprim was up to 64.65%;susceptibility rate to levofloxacin, azithromycin, ciprofloxacin, cefotaxime, chloramphenicol, cefuroxime, ampicillin / sulbactam was 97.96%, 96.84%,96.91%, 92.78%,85.71%,77.89% and 74.75% respectively. Of 100 H.influenzae isolates, 21 were βlactamase positive strains, and all were resistant to ampicillin. 35 ampicillinresistant strains were performed TEM and ROB genes detection, of which 22 were TEM positive, ROB positive strain was not found. ConclusionH.influenzae is sensitive to most antimicrobial agents except sulphamethoxazole/trimethoprim. Producing of TEM βlactamase is the major mechanism of ampcillin resistance in H.influenzae.