Simply supported bridges occupy the majority of bridges. Compared with the flexible large span bridges, Dynamic Amplification Factor (DAF) of them is relatively large and attracts lots of studies. However, most of these studies are focused on the simplified condition that a single vehicle passes through a single span bridge. In general, the bridge bears complex traffic flow rather than a single vehicle. The vehicle type and dynamic loads on the bridge are thus dispersed. It is necessary to explore the multi-vehicle effects on the DAF caused by the complex traffic flow. Also, adjacent spans of a multi-span simply supported beam bridge may have continuous dynamic effects on the vehicles. In this regard, the DAF of a simply supported bridge considering the multi-vehicle and multi-span effects are explored numerically based on the coupling dynamic analysis of traffic flow and bridge, which can also consider the acceleration and deceleration of the vehicles. Cellular Automata (CA) method is used to simulate the traffic flow. It is found that the worse the driving condition of the road roughness, the greater the DAF. When road roughness is Class C, the DAF exceeds the values of American and Chinese codes. Under the action of traffic flow, DAF of the first span of the multi-span simply supported beam is larger than that of the span under the single vehicle. Only considering the effect of a single vehicle may underestimate the dynamic impact on the bridge. Sparse traffic flow has a larger DAF than moderate traffic flow and dense traffic flow in the statistic meaning of the multi-span, because the driving speed of sparse traffic flow is closer to the resonant speed of the bridge and the vibration disharmony of multi-vehicles is smaller. The greater the deceleration of the traffic, the greater the dynamic impact on the bridge.