Aim: To investigate prevention measures of hospital-acquired infection and clinical features of suspicious COVID-19 patients in the pediatric respiratory ward. Methods: Patients with fever and cough or fever accompanied by vomiting and diarrhea were given pre-isolation measures. The medical records of the patients were collected and analyzed to summarize the pre-isolation measures and patients’ clinical features. Results: A total of 50 pre-isolated children with suspicious COVID-19 hospitalized between January 28 and March 5, 2020 were included in. Pre-isolation measures combined with epidemiological history and pathogen screenings were used to rule out COVID-19 patients. No definite COVID-19 cases were detected, while 2(4%) patients were suspected of having COVID-19. The pathogenic results mainly included Mycoplasma pneumonia (35, 70%).Thirty-five (70%) patients had bronchopneumonia. The pre-isolation patients had the common clinical epidemiological characteristics as patients with fever and cough, vomiting, and diarrhea. Discussion: Pre-isolation measures could prevent suspicious COVID-19 patients from coming in contact with other patients before definite exclusion. Clinical analysis of the patients was helpful for clinical nursing management. Conclusions: Pre-isolation measures combined with epidemiological history and pathogen screening (novel coronavirus nucleic acid and general pathogen) can help to minimize the risk of COVID-19 as a hospital-acquired infection.