In this study, the periodic seismic performance evaluation scheme is proposed using a structural health monitoring system in terms of seismic fragility. An instrumented highway bridge is used to demonstrate the evaluation procedure involving (1) measuring ambient vibration of a bridge under general vehicle loadings, (2) identifying modal parameters from the measured acceleration data by applying output-only modal identification method, (3) updating a preliminary finite element model (obtained from structural design drawings) with the identified modal parameters using real-coded genetic algorithm, (4) analyzing nonlinear response time histories of the structure under earthquake excitations, and finally (5) developing fragility curves represented by a log-normal distribution function using maximum likelihood estimation. It is found that the seismic fragility of a highway bridge can be updated using extracted modal parameters and can also be monitored further by utilizing the instrumented structural health monitoring system.