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The biofilm of Serratia marcescens are synthesized when it is linking to the surface within the aqueous environment. Some strains of S. marcescens are known to be problematic to the nosocomial infections because of its resistances to the drug. This paper aims to evaluate the antibiofilm efficacy of gold nanoparticles toward the clinical isolates of Serratia marcescens isolates. The anti-biofilm activity of AuNPs with subMIC concentration of (18.71) μg/ml was investigated using the microtiter plate method. The results exhibited that gold nanoparticles reduced the ability of Serratia marcescens to produce the biofilm by 51.6 – 80.4%.