Objective To analyze the bacterial types and drug resistance in patients with hepatolithiasis and biliary tract infection. Method The results of bile culture in patients with hepatolithiasis admitted from January 2008 to December 2017 were collected, and the difference in the types of drug-resistance bacteria and resistance rate between the patients of previous 5 years(group A)and the last 5 years(group B)were compared. Result In this study, 155 gram-negative bacteria(56.3%)and 120 gram-positive bacteria(56.3%)were detected. Enterococcus casselifavus was detected in 37 cases accounting for 13.3% of Gram-positive bacteria. The pathogens that accounted for the top 6 in isolation were Escherichia coli, Enterococcus faecalis, Enterococcus casselifavus, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus faecium. The detection rate of Gram-positive bacteria in Group A and Group B was 46% and 65%, respectively. Escherichia coli and Klebsiella pneumoniae had the lowest resistance rates to cefoperazone/sulbactam and imipenem. Cefoperazon/Schubatam and piperacillin/tazobartan were sensitive to Pseudomonas aeruginosa with a sensitive rate > 80%. Most of the antibiotics tested were sensitive to Enterococcus faecalis and Enterococcus casselifavus with a sensitive rate >90%, in which only one case with Enterococcus resisted to vancomycin. Conclusion Gram-negative bacteria still predominates in the biliary tract infection caused by hepatolithiasis, with an increasing detection rate of Gram-positive enterococci. The detection rate of Enterococcus casselifavus is high in this area and it has natural resistance to vancomycin, which needs more attention. Cefoperazon/Schubatan combined with ciprofloxacin/levofloxacin can cover the majority of bacteria detected in the bile of the patients with hepatolithiasis and biliary tract infection, which can be used as the preferred therapeutic regimen of experimental treatment.