This paper proposes a fatigue damage assessment methodology of a reinforced concrete bridge based on a train bridge coupled dynamic analysis system. This coupling system is composed of a vehicle a ballasted track and a bridge structure to realize a coupled vibration analysis during train passages. This methodology adopts the Palmgren Miner rule to linearly accumulate fatigue damage, and accomplishes fatigue assessment based on recommended SN relationships for reinforced concrete from various criteria. In this paper, the dynamic performances of a bridge structure are analyzed with two different vehicle models, a moving load model (and a moving spring mass damper model). Calculated dynamic stresses areused to evaluate the fatigue damage of critical positions in a reinforced concrete bridge. A case study is carried out of a railway arch bridge with a span of 89 m, Långforsen Bridge. It runs over Kalix River on the railway between Kalix and Morjärv in northern Sweden The fatigue assessment is made by combining the presented methodology with measurements. Furthermore, the effects of train speed and axle load on fatigue damage are investigated. The results indicate that good estimations of the fatigue damage assessment are made for four measured cases based on a train-bridge coupled dynamic analysis. For low speeds and light loads no difference could be seen in the cumulative fatigue damage for the two vehicle models. But high speeds and/or heavy load ssignificantly affect the fatigue damage.