The dynamic characteristics of bridge structures, such as the natural frequencies, mode shapes and model damping ratio, are the basis of structural dynamic computation, seismic analysis, vibration control and structural health condition monitoring. In this paper, a three-dimensional finite-element model is established for a highway bridge over a railway on No.312 National Highway and the ambient test is carried out in site, the dynamic characteristics of the bridge are studied using the finite-element analysis and ambient vibration measurements. Comparison between the theoretical and experimental results shows that the frequency differences of the modes range between 0.44% and 8.77%. If the measurement is more reliable, the finite element model updating is necessary. Thus, a set of de- sign variables is selected based on sensitivity analysis, then the finite ele- ment model of the bridge is updated based on optimization algorithm. The results of model updating show that the proposed updating method in this paper is more simple and effective, the updated finite element model can reflect the dynamic characteristics of the bridge better, the analytical results can provide the theoretical basis for damage identification and health con- dition monitoring of the bridge. The dynamic characteristics of bridge structures, such as the natural frequencies, mode shapes and model damping ratio, are the basis of structural dynamic computation, seismic analysis, vibration control and structural health condition monitoring. In this paper, a three-dimensional finite element model is established for a highway bridge over a railway on No. 312 National Highway and the ambient test is carried out in site, the dynamic characteristics of the bridge are studied using the finite-element analysis and ambient vibration measurements. Comparison between the theoretical and experimental results shows that the frequency differences of the modes range between 0.44% and 8.77%. If the measurement is more reliable, the finite element model updating is necessary. Thus, a set of de- sign variables is selected based on sensitivity analysis, then the finite ele- ment model of the bridge is updated based on optimization algorithm. The results of model updating show that the proposed up dating method in this paper is more simple and effective, the updated finite element model can reflect the dynamic characteristics of the bridge better, the analytical results can provide the theoretical basis for damage identification and health con- dition monitoring of the bridge.