Using truck weigh device, the data of traffic stream was collected from a toll station in Shaanxi province for 24 hours. A standard fatigue truck was developed based on the regular traffic data. Then S-N curves and Palgrem-Miner summation method was introduced in assessing fatigue life of the reinforcement in negative region of one continuous concrete bridge. It was concluded that the welding detail of reinforcement in negative region had long service life under normal traffic state. The fatigue life of reinforcement would be weakened considering overloading vehicles.