Introduction: The COVID-19 pandemic highlights the role of environmental cleaning in controlling infection transmission in hospitals. However, cleaning practice remains inadequate. An important component of effective cleaning is to obtain feedback on actual cleaning practice. This study aimed to evaluate the cleaning process quality from an implementation perspective. Methodology: An observational study was conducted in a tertiary public hospital in Wuhan, China and 92 cleaning processes of units housing patients with multidrug-resistant organism infections were recorded. The bed unit cleaning quality and floor cleaning quality were measured by six and five process indicators respectively. Descriptive statistics were used to describe the cleaning quality. Results: For bed unit cleaning quality, the appropriate rates of cleaning sequence, adherence to cleaning unit principle, use of cloth, use of cloth bucket, separation of clean and contaminated tools, and disinfectant concentration were 35.9%, 71.7%, 89.7%, 11.5%, 65.4%, and 48.7%, respectively. For floor cleaning quality, the appropriate rates of adherence to cleaning unit principle, use of cloth, use of cloth bucket, separation of clean and contaminated tools, and disinfectant concentration were 13.4%, 50.0%, 35.5%, 11.0%, and 36.7%, respectively. Conclusions: The cleaning staff showed poor environmental cleaning quality, especially the floor cleaning quality. The findings can help reveal deficiencies in cleaning practices, raise awareness of these deficiencies, and inform targeted strategies to improve cleaning quality and hospital safety.