The antimicrobial agents of silver nanoparticles (AgNPs) have been applied a little while back in diverse therapeutic studies. In this analysis, AgNPs were biosynthesized using an ecologically welcomed and cost-effective simple of bio-reduction. An isolate of Aspergillus flavus KF946095 (A. flavus) was found to biosynthesize AgNPs; the size of AgNPs was (56nm) and detected by UV-Vis analysis at (400 nm). The reducing properties for biosynthesis of AgNPs are mainly due to the protein functional surface reactive groups detected by Fourier Transform Infrared spectroscopy (FTIR).Whereas, FTIR for AgNPs showed different peaks at 3994.5, 3201.6, 1801.4, 1643.2 and 1604.7 cm^-1 that shared with the biosynthesize and stability of AgNPs as protein capping agents. Transmission Electron Microscope (TEM) confirmed the scattering of biosynthesized AgNPs within a sol with oval and round shapes. The antibiotic susceptibility test was studied for some pathogenic bacteria. Staphylococcus aureus DSM 1104 (S. aureus) appeared to be the more resistant strain; it resisted the action of 6 antibiotics out of 8 ones tested. MIC value of AgNPs was 20µg/mL and antibiotic ciprofloxacin was 30µg/mL. Mixture of MIC values or double MIC values distinctively inhibited the multidrug resistant (MDR) S.aureus.