Resistance spot welding (RSW) is the leading process of automobile body welding, and its quality is related to the reliability and safety of the whole vehicle. The effectively and automatically ultrasonic nondestructive testing is the development direction which can judge the spot welding quality, and then the positioning error of probe has an important impact on the test results. From this, conventional ultrasonic A-scan is adopted. Firstly, this paper presents the nugget diameter mathematic model of two-layer plates with different thickness when the probe positioning error is present. Then, the ultrasonic nondestructive testing simulation models of the spot welds are established, and the influence of the probe positioning error on the calculation result of the nugget diameter is studied. Finally, this paper carries out ultrasonic nondestructive testing of spot welding of two-layer plates with different thickness when the probe has positioning error. The experimental results verify the correctness of the theoretical analysis and simulation.