Abstract: As a common biliary disease, cholelithiasis has high incidence and recurrence rates, with negative impact on patients′ health and quality of life. The causes of cholelithiasis are complex, and recent studies have shown that microorganisms may be associated with the formation and recurrence of cholelithiasis, but the specific relationship between the two has not been fully elucidated. Bile microbiota may be related to the increase of incidence rate of gallstones. Different types of cholelithiasis have different microbial composition. Microorganisms may participate in the formation and recurrence of gallstones by influencing bile composition, promoting the formation of stone cores and other ways. The authors provide a review on the etiology and pathogenesis of cholelithiasis, classification of cholelithiasis, microenvironment of biliary microbiota, and the relationship between biliary microbiota and stone formation, aiming to provide new ideas and theoretical basis for the prevention and treatment of cholelithiasis.