ObjectiveTo investigate the distribution and antimicrobial  resistance of clinical isolates of Haemophilus influenzae (H. influenzae) from Hubei province.MethodsClinical isolates of H. influenzae were collected from 15 tertiary hospitals in Hubei province  from 2008 to 2010. Antimicrobial susceptibility testing was performed  by KirbyBauer disc diffusion method and results were analyzed according to CLSI 2009 guideline;βlactamase was detected with nitrocefin disk testing.ResultsA total of 855 H. influenzae isolates were collected, 673(78.71%) of which were from inpatients ; 87.37% (747 isolates) were from respiratory samples; 575 isolates (67.25%) were  from adults , and 280 (32.75%) from children,56.07% (157 isolates) of which were  from children under  2 years old . The resistant rate to trimethoprimsulfamethoxazole,ampicillin,chloramphenicol and ampicillin/sulbactam was  58.0%,42.9%,16.9% and 16.2%, respectively, the resistant rate to   cefuroxime, cefotaxime, ciprofloxacin, levofloxacin  and azithromycin were all <10%. 232 isolates (27.13%) produced betalactamase; 22.26% of adult isolates and  37.14% of child isolates  were betalactamase positive . A total of 32 isolates were  betalactamase negative ampicillinresistant (BLNAR) H. influenzae.ConclusionMost  isolated H. influenzae from  Hubei province were from respiratory samples of inpatients,the resistance  to trimethoprim/sulfamethoxazole and ampicillin are high, the prevalence of betalactamase isolates from children is higher than that from  adults. The primary mechanism of ampicillin resistance in H. influenzae is production of betalactamase.