Objective To investigate the distribution and antibiotic resistance of bacterial isolates from blood samples in pediatric patients in tertiary hospitals in Hubei area from 2006 to 2007.Methods Pathogenic bacteria isolated from blood samples of pediatric patients were collected from 17 tertiary hospitals in Hubei area from 2006 to 2007. All strains were isolated and identified by routine Methods . Antimicrobial susceptibility testing was conducted on all isolates using Kirby-Bauer Methods . Results A total of 941 strains were collected from January to December of 2006. The most common microorganism was coagulase-negative Staphylococcus (573, 60.9%), followed by Staphylococcus aureus (127, 13.5%), Enterococcus faecalis (33, 3.5%), Escherichia coli (16, 1.7%). A total of 969 strains were collected from January to December of 2007. The most common species was coagulase negative Staphylococcus (583, 60.2%), followed by S. aureus (162, 16.7%), E. faecalis (28, 2.9%), E. coli (21, 2.2%), E. faecium (11, 1.1%), Salmonella choleraesuis (11, 1.1%). Of the isolates collected during 2006, the prevalence of methicillin-resistant S. aureus (MRSA) was 71.7%. The prevalence of ESBLs was 56.2% in E. coli. Of the isolates collected during 2007, the prevalence of MRSA was 79.6%. The prevalence of ESBLs was 47.6% in E. coli. MRSA strains were more resistant to antibiotics than methicillin-susceptible S. aureus (MSSA). No glycopeptide-resistant strain was identified in Staphylococcus. Conclusions Staphylococcus is the most frequently isolated pathogen from blood samples of pediatric patients in tertiary hospitals in Hubei area.