Objective To evaluate the clinical applicability of clinical practice guidelines in China in the past 5 years, and to provide suggestions for better development and implementation of guidelines. Methods A cross-sectional study was conducted. A total of 61 guidelines issued by the Chinese Medical Association and the Chinese Medical Doctor Association from 2015 to 2020 were selected randomly based on CNKI database. Each guideline was evaluated by medical staff from three regions using the instrument for evaluating clinical applicability of guidelines (version 2.0). Quantitative data were described by mean, standard deviation, median and interquartile range, while qualitative data were described by frequency and composition ratio. Results A total of 6 904 valid questionnaires were collected from 119 medical institutions in 26 provinces of China. The results showed that the standardized scores ranged from 54.4% to 72.7% (median 64.3%) for accessibility, from 66.1% to 83.3% (median 74.9%) for readability, from 53.7% to 94.7% (median 86.0%) for acceptability, and ranged from 61.4% to 81.0% (median 72.5%) for feasibility. Approximately 4.3% of the medical staff considered that there were obstacles to implement the guidelines, including environmental factors (32.1%), medical staff factors (26.5%), guidelines factors (22.5%), and patients or family factors (18.9%). Conclusion The overall clinical applicability of the guidelines selected in this study is satisfactory. However, it is necessary to improve the accessibility of clinical guidelines. In addition, to improve the clinical applicability, it is suggested that medical staff should be trained and provided with supporting resources.