In the present paper, pure and silver-doped zinc oxide (Ag-doped ZnO) nanoparticles were prepared using a green route and employing Salvadora persica leaf extract. Antibacterial performance of synthesized nanoparticles (NPs) survey on gram-negative and gram-positive bacteria through disk diffusion and micro-dilution technics was also studied. The synthesized NPs were identified by X-ray diffraction (XRD), ultraviolet–visible (UV–Vis), field emission scanning electron microscope (FESEM), transmission electron microscopy (TEM), and energy-dispersive X-ray (EDX) analyses. XRD and EDX results presented that the silver was well doped into the structure of ZnO. In addition, spherical shape of this product, as well as the average size of 25–40 nm, were observed in the results of FESEM and TEM images. The results of disk diffusion and microdilution of synthesized nanoparticles showed that Ag-doped ZnO nanoparticles, have more antibacterial performance than pure zinc oxide nanoparticles on Staphylococcus aureus and Escherichia coli. So, synthesized NPs can be proposed as a choice in clinical and laboratory applications.