The review article provides definitions and presents current data on the biological significance of the microbiome and microbiota of urine. The urinary microbiota is less well understood than the intestinal microbiota, but its biological role is complex and multifaceted. To date, its importance in the formation of colonization resistance, which prevents the invasion of uropathogens, is obvious. An analysis of methods for assessing the microbiome and microbiota of urine was carried out. The shortcomings of widely used standard microbiological research methods are shown. The method of 16S rRNA gene sequencing is described in detail. Information about the gender characteristics of the urine microbiota and the results of its study in healthy women and men are given. Further progress in the study of the urobiome is associated with the acquisition of new technologies for genomic research and the development of bioinformatics.