The invention discloses a development culture medium for separating and identifying pathogens in a urogenital tract, which at least comprises three development substrates, namely a hexosaminidase substrate, a beta-D-galactosidase substrate, a beta-D-glucuroide substrate. The development culture medium added with the three development substrates is prepared into a microbial culture medium agar plate, a sample or a bacterial colony subjected to separate culture is inoculated into the development plate and is incubated, and a result can be directly observed. In the development culture medium, various pathogens including bacteria and fungi in a genital tract can be simultaneously cultured and identified on the same development plate, namely bacterial pathogens such as Escherichia coli, Klebsiella pneumonia, enterococcus, pseudomonas aeruginosa, proteus mirabilis, staphylococcus aureus and the like and fungal pathogens such as candida albicans, candida tropicalis and the like can be simultaneously identified, and judgment can be performed through visual inspection; and the development culture medium is quick and convenient to use, and easy to operate.