The high concentration of viral bioaerosols within the negative pressure isolation wards could pose a challenge to preventing potential cross-infection amongst healthcare workers (HCWs) and patients. Using the Euler–Lagrange methodology, this study numerically simulated the spatial and temporal distribution characteristics of bioaerosols in a typical negative pressure isolation ward as well as determined the interaction of ventilation mode and patient posture on ward ventilation performance. The removal effect of particle groups produced by two respiratory behaviours (breathing and coughing) was quantitatively analyzed, and the effect of exhaust air ratio and air exchange rate on particle distribution was discussed. The results showed that the migration characteristics of bioaerosol particles were sensitive to both the ventilation pattern and patient posture, which showed significant interactions. On this basis, the ventilation pattern with the best ventilation performance was evaluated, showing a particle removal effect of 70–85%. Due to the initial momentum difference, the diffusion behaviour of cough and breath particles was not consistent, but optimizing the airflow distribution near the exhaust outlet could improve their removal efficiency in the meantime. Further studies found that equal exhaust air velocity ratio facilitated the removal of aerosol particles, and an appropriate increase in the air exchange rate could also reduce the particle content.