The invention discloses a biphasic media for genitourinary tract mycoplasma, and belongs to the field of microbiological detection. The biphasic media comprises a liquid medium and a solid medium, wherein the solid medium is arranged in a container and forms an inclined plane; meanwhile 1/4 to 1/2 of the inclined plane is immersed into the liquid medium; the biphasic media mainly comprises the following components: yeast extract powder, beef heart infusion, soy peptone, sodium chloride, dipotassium phosphate, manganese sulfate, cysteine hydrochloride, urea, arginine, glutamine, equinum serum,a DMEM liquid medium, sodium pyruvate, a phenol red indicator, nysfungin, penicillin and agar; the container further comprises a sealing component capable of sealing the inner space of the container and a filter screen; a carbon dioxide producing piece is arranged in the filter screen, which can absorb moisture during culturing and release carbon dioxide, so that carbon dioxide incubators are notrequired any more. The whole process is simple to operate, the specificity is high, and laboratory operation is easy.