ObjectiveTo evaluate detection effect of three methods on monitoring microbes in dialysate and dialysis water for hemodialysis. MethodsSeventytwo dialysate and dialysis water specimens were collected from 36 medical institutions, specimens were cultured with three methods: blood agar plate incubated at 35℃ for 72 hours, Tryptic soy agar（TSA）plate incubated at 35℃ for 72 hours, and Reasoner’s 2A agar (R2A agar) plate incubated at 23℃ for 168 hours, colony counts, isolation of colony, and detection rate of colony exceeding action level（≥50 CFU/mL） were compared among three methods. ResultsThe colony isolation rates of microbes in dialysate and dialysis water detected by blood agar plate, TSA plate and R2A plate were 40.28%, 63.89%, and 69.44% respectively, difference was significant（χ2=14.16，P＜0.05）；pairwise comparison showed that isolation rates of colony on R2A agar plate and TSA plate were higher than blood agar plate. There was significant difference in isolated colony count between blood agar plate and R2A agar plate, TSA plate and R2A agar plate respectively（Z=-4.515, -6.970 respectively，both P＜0.05）. The rates of isolated colony exceeding action level in dialysate and dialysis water detected by blood agar plate, TSA plate, and R2A agar plate were 1.39%, 4.17%, and 20.83% respectively, difference was significant（χ2=19.83，P＜0.05），detection rate of R2A agar plate was higher than the other two methods. ConclusionThe detection rate of colony by R2A agar plate and TSA plate are better than blood agar plate, detection rate of colony exceeding action level by R2A agar plate is higher than TSA plate and blood agar plate, R2A agar plate for microbial monitoring(23℃,168 h) on dialysate and dialysis water is superior to the other two methods.