Nanoparticle biosynthesis has received increasing attention due to the growing need to develop safe, cost-effective and environmentally friendly technologies biosynthesized for nanomaterials. Therefore, this study aimed to study the effect of silver nanoparticles manufactured from Penicillium digitatum  , against bacteria and measure the level of interference of antibiotics with nanoparticles. In this paper, silver nanoparticles (Ag NPs) were synthesized using the reduction of the Ag + aqueous ion from Penicillium digitatium.The nanoparticles of Ag NPs were also tested by and X-ray diffraction and transmission electron microscope (TEM) and UV-visible light absorption spectroscopy method and field-emission scanning electron microscopy (FESEM). Samples is 70 were collected from patients visiting Samarra General Hospital in Samarra city during the period from December 2021 to May 2022 and of both sexes with ages ranging from 1 to 60 years. Samples were collected from different sources of infections, The inhibitory ability of silver nanoparticles produced from mushrooms was studied on pathogenic bacteria, and that was done after making four concentrations, namely 10%, 25%, 50%, and 100% with the control without any concentration of the nanomaterials. The inhibitory activity of silver nanoparticles Ag NPs was tested on some pathogenic bacterial species isolated from burns, wounds, urinary tract and intestinal infections  . After that, biochemical tests were performed to diagnose the bacteria, such as the catalase test, the oxidase test, and the urease test, in addition to microscopic examination to confirm the diagnosis by the well diffusion after the sensitivity test was measured. The results showed an inhibitory efficacy of processed silver particles at a concentration of 25, 50, 75 and 100% towards clinical samples for resistance to pathological bacteria Proteus mirabilis, Acinetobacter baumannii. The role of silver nanoparticles is evident in inhibiting the growth of antibiotic-resistant bacteria, where biologically produced silver nanoparticles can be used as alternatives to antibiotics as the production of nanomaterials is inexpensive and biomaterials are available.