This prospective, three-centre study tested for antimicrobial susceptibility in 898 isolates of Haemophilus influenzae between 2000 and 2002 in Chinese children aged under 5 years with acute upper respiratory tract infection. The average incidence of P-lactamase production was 12.0%. Overall, 88.0% of isolates were susceptible to ampicillin, 100.0% were susceptible to arnoxicillin/clavulanic acid, ceftriaxone, cefuroxime and azithromycin, and 99.0% were susceptible to ciprofloxacin. Isolates from Beijing and Shanghai had a lower susceptibility to tetracycline (57.0% and 61.0%, respectively) compared with those from Guangzhou (81.0%), while trimethoprim/sulfamethoxazole susceptibilities in Shanghai (47.0%) and Guangzhou (54.0%) were significantly higher than in Beijing (35.0%). A total of 34.5% of all the isolates were susceptible to all eight of these antimicrobial agents and 12.8% were multi-drug resistant. Ampicillin resistance increased over the duration of the study. These findings show that P-lactamase production and ampicillin resistance among isolates from Chinese children with upper respiratory tract infection are increasing, and highlight the strong correlation between ampicillin resistance and resistance to cefaclor, chloramphenicol and tetracycline in H. influenzae isolates.