Buildings of strategic importance should be able to resist seismic forces in accordance with potential earthquakes that may occur at the location and remain fully operational afterwards. However, many of them were constructed before the modern principles of seismic design were known (especially regarding detailing and ductility), and therefore may be considered substandard. The first step in mitigating the seismic risk of such structures is to assess their seismic performance and, in particular, to identify their structural deficiencies. This study presents a comprehensive methodology for the seismic performance assessment of individual buildings, applied to an existing reinforced concrete (RC) hospital. This building is of an irregular layout, constructed as a structural wall system, and it is located in the seismically active region of Croatia. It includes the assessment of seismic hazards on the location, ambient noise measurements, experimental determination of structural modal parameters, creation of a detailed numerical model calibrated with experimental data, and a seismic performance assessment using various analysis methods. As a result, the building collapse mechanisms were determined and critical structural elements identified, which is the basis for future actions directed to the reduction of its risk (e.g., applications of specific measures for a target retrofit, proposal of evacuation routes and safe places inside the building, etc.).