Objective To evaluate the accuracy of fluorescent labeling method and microbiological method in monitoring the quality of clean and hygiene of high frequency touched object surface environment in intensive care unit (ICU). Methods In the last month of each quarter from the third quarter of 2017 to the second quarter of 2019, fluorescence labeling and microbiological method was used to monitor and sample surface of high frequency touched object surface in general ICU (GICU) and respiratory ICU (RICU), qualified rate of surveillance of two methods in the same period was compared. Results From the third quarter of 2017 to the second quarter of 2019, a total of 8 times of surveillance were carried out, results showed that qualified rates of fluorescence labeling clearance in RICU and GICU were 41.62% and 71.45% respectively. Qualified rates of microbiological method in RICU and GICU were 45.58% and 45.18% respectively. Qualified rate of fluorescence labeling method in RICU increased gradually, the lowest was in the third quarter of 2017, which was 28.42% lower than that of microbiological method, and the highest was in the first quarter of 2019, which was 24.90% higher than that of microbiological method; qualified rates of fluorescence labeling method in GICU were all higher than microbiological monitoring method, difference of qualified rates of two methods were 2.86%-44.66%. Conclusion Fluorescence labeling method can be used as the evaluation standard of environmental hygiene and cleaning quality, but it needs to be combined with microbiological monitoring method to achieve more accurate and meaningful result.