During the last decades orthotropic steel decks have become a widely used component of steel bridges, however high numbers of fatigue cracks have been recently detected in these structure types. It is well known, that the orthotropic decks are sensitive to fatigue damage, therefore safe fatigue design is important for the safety assessment of bridge structures. The fatigue damage assessment and life prediction of steel bridges can be carried out by using online structural health monitoring systems (SHM). By using the SHM system, it is possible to detect deterioration, determine anomalies and assess the safety level which will allow optimum maintenance strategies. On-structure long-term monitoring systems have been already successfully implemented on bridges all around the world in order to detect cracks just like on the orthotropic steel deck structure of M0 Háros Danube highway Bridge in Hungary. The purpose of the current research program is the investigation and analysis of the fatigue behaviour of this orthotropic steel bridge deck and the evaluation process improvement of the data rows collected by the monitoring system. Additional laboratory tests are carried out at 6 large scale test specimens at the Budapest University of Technology and Economics, Department of Structural Engineering to determine the fatigue resistance of closed section longitudinal stiffeners. In parallel finite element model is developed to simulate the fatigue behaviour of the analyzed components and to investigate its fatigue behaviour by numerical analysis. The aim of the current study is to analyze and verify the structural behaviour of the orthotropic steel deck on the basis of the executed test results, numerical simulations and the strain-time history data from the online structural health monitoring system implemented on the studied bridge.