Objective To describe the changing trend and related factors of prevalence rates of healthcare-associa-ted infection (HAI) in a tertiary hospital in the past 10 years, and analyze the influencing factors for HAI. Methods A cross-sectional survey on HAI was conducted for 10 consecutive years from 2013 to 2022 (one day was selected as the survey day each year), data were collected. The distribution and related factors of prevalence rates of HAI were analyzed by trend-χ² test and Pearson correlation coefficient. Multivariate logistic regression and multilayer perceptron (MLP) models were constructed to analyze the independent effect and significance of factors. Results From 2013 to 2022, the prevalence rates of HAI ranged from 4.66% to 8.07% in this hospital, showing a linear upward trend. The proportions of ICU patients and utilization rate of central venous catheters within 2 days before the survey showed linear upward trends, while the proportion of patients with urinary catheters within 2 days before the survey and proportion of patients undergoing surgery within 30 days before the survey decreased. The MLP model revealed that the top 3 important factors for HAI were length of hospital stay >10 days, admission in ICU, and indwelling central venous catheters within 2 days before the survey. Multivariate logistic regression model indicated that length of hospital stay ＞10 days, indwelling central venous catheters or urinary catheters within 2 days before the survey, surgery within 30 days before the survey, and admission in ICU were independent influencing factors for HAI. Conclusion The incidence of HAI in this hospital presents a linear increase in recent 10 years, the causes should be further analyzed and the direction of intervention should be determined through targeted surveillance. Adopting trend test statistical analysis method, logistic regression, MLP multi-factor model can further explore the data value of HAI prevalence survey.