Micro defects, such as pore and inclusions, inevitably appear in the forming process of three-dimensional (3D) printed materials, which would affect the mechanical properties of materials. Therefore, a nondestructive testing method is urgently needed to evaluate the effect of these micro defects on the mechanical properties of materials. In the present work, by using a nonlinear ultrasonic testing technology, mechanical test and characterization of material microstructure, the relationship between the relative acoustic nonlinearity parameter (RANP), tensile strength and material defect ratio of 3D printed aluminum alloy specimens under different scanning powers is investigated. The analysis results show that the greater the material defect ratio is, the smaller the tensile strength is and the greater the RANP is, and the RANP could be used to evaluate the strength of materials. Moreover, fatigue damage induced by high cycle fatigue loading test in the first stage of early performance degradation, the results of nonlinear ultrasonic testing show that the RANP presents an increasing trend as the fatigue load increases. By observing changes in material microstructure, it is found that the increasing acoustic nonlinearity parameter is due to the directional coarsening degree of the precipitated phase increasing, which shows that RANP is very sensitive to the change of material microstructure. The above results show that, the nonlinear ultrasonic testing technology can quantitatively evaluate and predict the mechanical performance and early performance degradation of 3D printing aluminum alloy.