Abstract：Objective Adenosine triphosphate (ATP) bioluminescence and microbiological culture methods were used for evaluation of cleaning procedure changed before or after in hemodialysis room. These provide a real-time and effective method on monitoring and evaluation of the environmental clean level of hemodialysis room. Methods Choose five sites from the hemodiaylsis room and use ATP bioluminescence and microbiological culture methods to monitor and record the clean level of those sites before and after changing clean procedure, comparison of two results and evaluation of a new clean procedure. Results The mean of ATP bioluminescence method before changing clean procedure respectively is 788. 310. 530. 600 and 900RLU, after cleaning procedure changed is 176. 130. 120. 220 and 239RLU.The mean of microbiological culture method respectively is 12. 4. 9. 6 and 8cfu, after cleaning procedure changed is 2. 1. 1. 1 and 2cfu. The surface contaminated level of homedialysis room was significantly reduced, the monitoring results of ATP bioluminescence are similar with microbiological culture method. Conclusions Effectively surface cleaning procedure can dramatically reduced the surface contaminated condition, ATP bioluminescence is much more quick and efficient technology used in monitoring and evaluation on the nosocomial environmental clean level.