ObjectiveTo realize isolation state and drug resistance of Escherichia coli (E.coli) from  hospital environment, clinical samples and health care workers(HCWs). MethodsProspective investigation was performed. From May 1, 2008 to October 31,2008, E.coli were isolated from samples of environment of  intensive care unit (ICU), ICU HCWs and nonICU HCWs at a large general hospital. Data of E.coli isolated from patients were from clinical microbial laboratory during the same period. Drug resistance of E.coli were analyzed statistically.ResultsA total of 376 samples were collected  from general  ICU environment, 69(18.35%) strains of E. coli were isolated. The isolation rate of E.coli from handwashing facilities, sputum aspirators and respirators was 29.17%, 28.07% and 21.74% respectively. The isolation rate of E.coli from 177 HCWs’ anal swabs was 45.76%(81/177). 73 strains of E.coli were isolated from clinical samples of patients, most from stool (21 strains, 28.76%) and urine (18 strains, 24.66%) samples. Except resistant rate of clinical samples to cefoperazone/sulbactam (13.70%) was lower than 30.43% of environmental samples (χ2=5.82, P<0.05), the differences in drug resistance  to the other antimicrobial agents were not significant (χ2=0.56-2.89, P>0.05); There were no significant difference in drug resistant rates of E. coli from HCWs, clinical  and environmental samples to gentamycin, nitrofurantoin and ceftriaxone (χ2=1.34-3.12,P>0.05), but resistant rates of  E. coli from environmental and clinical samples were higher than  that of  HCWs (χ2=10.59-215.28, P<0.05); the difference in drug resistant rates of E. coli from HCWs in ICU and nonICU was not significant (χ2=0.18-2.36,P>0.05). ConclusionDrug resistance of E. coli from environment is closely related with patients, E. coli from environment and  patients’ samples has little effect on drug resistance of intestinal E. coli from HCWs.