A negatively-pressurized isolation ward must maintain a ward pressure lower the ambient pressure. which prevent the pathogen from escaping into the environment. The ventilation in the ward is also required to remove the pathogen effectively. This study measured the effective air change rate at different locations in a negatively-pressurized ward by the concentration decay of trace gas concentration. The resident times were also measured with trace gas released at different locations in the ward. The time for 50% of trace gas being exhausted, t50%, was used as an index. The results of air change rate and t50% was compared at nine locations in the ward. The ventilation dead space and anomaly airflow, both would prevent the pathogen to be effectively removed, were able to be identified by the experimental results. It was found that the airflow generated by an air-condition in the ward may also have a significant effect on the pathogen-removing.