Abstract: Objective To explore the distribution and drug resistance of pathogenic bacteria in blood culture microorganism test, so as to provide reference for effective control of the occurrence and spread of blood stream infection. Methods A total of 4 790 inpatients in Yuhuan People's Hospital from July 2018 to June 2019 were selected, 4 790 blood samples were collected and sent for examination. BacT/Alert was used 3 D type automatic blood culture instrument and its supporting culture bottle were used for blood culture, and positive strains were smeared and transferred in time. Vitek-2 compact automatic microbial analysis system was used for strain identification and drug sensitivity test, and the isolated bacterial strains and drug resistance were analyzed. Results In 4 790 blood samples, 523 pathogenic bacteria were isolated, the positive rate was 10.92%, including 283 gram-negative bacteria(54.11%), 230 gram-positive bacteria(43.98%), 8 fungi(1.53%), and 2 anaerobes(0.38%). The distribution of pathogenic bacteria was mainly in ICU(25.91%), general surgery(22.75%), and infection department(13.77%). The drug resistance of gram-negative bacteria and gram-positive bacteria were significantly different. The resistance of Escherichia coli and Klebsiella pneumoniae to ampicillin was high(>82.00%), to imipenem was low(<5.00%). The resistance of Acinetobacter baumannii to furantoin, ampicillin, aztreonam, and cefazolin were high(100.00%), and to tobramycin was low(27.27%). The resistance of gram-positive bacteria to erythromycin and penicillin were generally high(>76.00%), but not resistant to linezolid and vancomycin(all 0.00%). Conclusion There are many kinds of pathogenic strains in the blood culture microorganism test, the departments are widely distributed, and the situation of drug resistance is severe. Clinicians should pay more attention to the results of early detection and drug resistance analysis of pathogenic strains in blood culture microorganism test, so as to improve clinical diagnosis and application of antibiotics.