Abstract: OBJECTIVE To investigate the test result of samples from clinical microbiological laboratory, explore the distribution and drug resistance of pathogens, so as to provide evidence for clinical prevention and treatment. METHODS The data of test result of samples from Apr. 2014 to Oct. 2015 in clinical microbiological laboratory were retrospectively analyzed. The sources and positive rate of samples, and the distribution and drug resistance of pathogens were investigated and analyzed. RESULTS Totally 2784 strains of pathogens were isolated, including 2365 strains of gram-negative bacteria, accounted for 84.9%, 344 strains of gram-positive bacteria, accounted for 12.4%, and 75 strains of fungi, accounted for 2.7%, respectively. The gram-negative bacteria included Acinetobacter baumanii, Escherichia coli, Pseudomonas aeruginosa, etc, and they were resistant to ampicillin, compound sulfamethoxazole, with the drug resistant rate ＞50%. The gram-positive bacteria including: Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, etc, and they were resistant to penicillin G, with the drug resistant rate ＞50%. CONCLUSION Most of pathogens are gram-negative bacteria which were multi-drug resistant. It should be paid attention to hospital management.