The ocular surface microbiome is a microbial community composed of commensal and pathogenic microorganisms, which plays a crucial role in maintaining the homeostasis of the ocular surface microenvironment. Dysbiosis of ocular surface microbiota is not only involved in the occurrence and progression of ocular surface diseases such as keratitis, conjunctivitis, and dry eye disease (DED), but also related to dry eye complications caused by various systemic immune diseases. DED is an ocular surface disease related to multiple factors, and its prevention and treatment have become an important public health issue due to changes in lifestyle and population aging. DED is closely related to the imbalance of ocular surface microbiota. The commensal microbiota on the ocular surface can influence the occurrence and progression of DED through mechanisms such as immune tolerance and inflammatory responses, and it also contributes to the vicious cycle of DED complications, but its specific pathways remain unclear. This review explores the correlation between ocular surface microbiota and DED, including differences in the composition of ocular surface microbiota between healthy individuals and DED patients, the correlation between the two, as well as the applications of ocular surface microbiota in the diagnosis and treatment of DED, aiming to provide new insights for its prevention and management.