Abstract: Objective To analyze the bacteria distribution and drug resistance in our hospital from 2011 to 2013, and provide basis for clinical anti-infection treatment. Methods The isolation rate, distribution and drug resistance of bacteria and fungi isolated in clinical specimens of our hospital were retrospectively analyzed. Results In the isolated bacteria collected from patients in our hospital during the past three years, gram-negative bacteria accounted for the highest isolation rates, which were 55.0%, 57.6% and 59.8%, respectively. Gram-positive bacteria showed stable isolation rates, which were 23.3%, 22.9% and 22.1%, respectively. Fungi's isolation rates reduced year by year. Sputum, urine and blood were the top three isolation rates in the specimen sources in the last three years. Pseudomonas aeruginosa, E. coli, candida albicans, klebsiella pneumoniae and acinetobacter baumannii were the most common seen isolated bacteria. E. coli and klebsiella pneumoniae were sensitive to imipenem and meropenem, and the resistance rate was less than 10%. Acinetobacter baumannii and pseudomonas aeruginosa were sensitive to polymyxin B and amikacin with the resistance rate of more than 60%. Staphylococcus aureus which had resistance to teicoplanin and vancomycin was not found. Respiratory fungi had high separation rate, which was mainly showed in candida albicans. The sensitivity of amphotericin B, fluorine cytosine and voriconazole were good, which was more than 90%. Conclusion Gram-negative bacteria are the dominant bacteria during the past three years, and the pathogen mainly comes from sputum. Imipenem and meropenem are sensitive to E. coli and klebsiella pneumonia bacteria. Polymyxin B and amikacin are sensitive to acinetobacter baumannii and pseudomonas aeruginosa. Hospital cross infection can be reduced by strengthening the monitoring of bacterial drug resistance and using antibacterial agents reasonably.