Objective To understand the occurrence of healthcare-associated infection (HAI) in patients with coronavirus disease 2019 (COVID-19), and provide basis for formulating prevention and control measures during the epidemic period. Methods By using point prevalence survey and combining bedside survey with medical record survey, occurrence of HAI, antimicrobial use and invasive manipulation of all hospitalized patients with COVID-19 in a hospital on March 24, 2020 were investigated. Results A total of 323 patients with COVID-19 were investigated, with an average age of (58.33±14.30) years old; the average length of hospitalization stay was (13.13±7.23) days. Prevalence rate of HAI was 3.10%, and that of critically ill patients was the highest (33.33%); the main infection site was lower respiratory tract (30.00%); incidence of catheter-associated urinary tract infection, central line-associated bloodstream infection and ventilator-associated pneumonia were 5.88%, 7.69% and 0 respectively; antimicrobial use rate was 33.44%, 82.41% of which were prophylactic antimicrobial use; 84.26% of which were monotherapy and 68.52% of which were oral administration; the most frequently used antimicrobial agents was moxifloxacin (23.84%). Conclusion The prevalence rate of HAI in patients with COVID-19 is slightly higher than the national average level in 2014, special attention should be paid to the prevention and control of secondary pulmonary bacterial infection in critically ill patients, and routine use of antimicrobial agents is not recommended to prevent bacterial infection.