Microbial antibiotic resistance is rapidly increasing as a result of overuse or misuse of antibiotics, as well as a lack of new, effective antibiotics. Alternative antimicrobial treatments, such as nanoparticles, and their potential for stronger synergetic effect when paired with other active chemicals, could be a viable option. This study is prepared to estimate the antibacterial activity of silver nanoparticles (AgNPs) that have been synthesized using the biomass-free filtrate of Aspergillus flavus. The formation of AgNPs was reported by color changed to a dark brownish-black after 72 hours of incubation. The AgNPs surface plasmon resonance peak was indicated in the UV–Vis spectrum at 427 nm. The synthesis of AgNPs with a nanoparticle size of 10 to 35 nm was validated using transmission electron microscopy. The increase in folding area was calculated to detect the synergistic potential of the combined AgNPs with a broad range of conventional antibiotics. AgNPs have broad-spectrum activity against all strains tested. The most sensitive strain was Escherichia coli (11 mm), whereas the most resistant strain was Pseudomonas aeruginosa, as indicated by the lowest inhibition zone (7 mm). The lowest Minimum Inhibitory Concentration indicated was against K. pneumonia and Enterobacter cloacae (0.025 mg/mL, each), followed by Staphylococcus epidermidis (0.05 mg/mL), E. coli and Shigella sp. (0.075 mg/mL, each), and then S. aureus (0.1 mg/mL). Notable synergy was reported between AgNPs and either ampicillin, erythromycin, ceftriaxone, vancomycin, azlocillin, or amoxicillin against S. aureus in the range between 29.3-fold to 8-fold. In addition, synergy was seen between AgNPs and either vancomycin, clindamycin, or erythromycin against P. aeruginosa (31.1-8.0-fold). Also, a maximum increase in IFA when erythromycin and vancomycin were synergized with AgNPs against E. cloacae was reported (IFA of 10.0 and 9.0, respectively). Similarly, AgNPs with either aztreonam or azlocillin against E. coli and amoxicillin, ciprofloxacin, or ceftriaxone against Shigella sp. caused an increase in the fold area of inhibition of between 5.3-3.7-fold. This result may have an advantage in encouraging the use of combined AgNPS with conventional antibiotics in treating infectious diseases caused by antibiotic-resistant bacteria.