In view of the non-destructive and non-contact features, laser ultrasonic (LU) technology has long been the effective method to detect tiny defects for laser powder bed fusion (LPBF) additive manufactured specimens. Of larger concern is the variation and the corresponding mechanism on tested results of LU detection as the property of LPBF additive manufactured specimen is changed. Aiming at the property of surface roughness, this work investigated the propagation characteristics of excited ultrasonic waves in LPBF additive manufactured 316L stainless steel with different surface roughness, as well as the interaction between ultrasonic waves and artificial submillimeter holes. Both numerical simulated and experimental study were conducted. Simulated results revealed that the amplitudes of longitudinal wave (L wave) and its echo wave L1 at the holes exhibited a discernible increase as the surface was coarser. The increase in surface roughness was detrimental to the resolution of defect detection as was expected from the increased amount of noise. LPBF fabrication and the subsequent LU pulse-echo detection were conducted for 316L stainless steel. Both B-scan and C-scan were able to detect the holes with the diameter of 0.6 mm. The speckle phenomenon deriving from the increase in surface roughness emerged, corresponding to the increased ultrasonic signal energy but deteriorated resolution of detected images. It is feasible to optimize LU detected effect by minimize the surface roughness of tested specimens.