Life-cycle cost analysis (LCCA) is a decision-making tool particularly useful for the design of bridges as it predicts lifetime expenses and supports the inspections management and the maintenance activities. LCCA allows to consider uncertainties on loads, resistances, degradation and on the numerical modelling and structural response analysis. It also permits to consider different limit states and different types of damage in a unified framework. Among the types of damages that can occur to steel and steel-concrete composite bridges, fatigue is one of the most dangerous ones, as it may lead to sudden and fragile rupture, even at operational traffic levels. In this context, the present paper proposes a framework for LCCA based on the use of the Pacific Earthquake Engineering Research (PEER) equation which is for the first time utilized for fragility and cost analysis of bridges subjected to fatigue, highlighting the possibility of treating the problem of fatigue damage estimation with an approach similar to the one currently adopted for damage induced by other hazards, like earthquake and wind. To this aim, a damage index computed through the Palmgren-Miner’s rule is adopted as engineering demand parameter. The framework is applied to a composite steel-reinforced concrete multi-span roadway bridge by evaluating the fatigue limit state from different traffic load models, i.e. a Technical Code-based model and a model based on results of Weigh in Motion monitoring system. The evolution over time of the probability of failure and the life-cycle costs due to fatigue damage induced by heavy traffic loads are investigated for different probability distributions of the engineering demand parameter and for different fragility models. The comparison between the fatigue failure probabilities and the life-cycle costs obtained with the two traffic models, encourages the adoption of traffic monitoring systems for a correct damage estimation.