The synthesis of (+)AuNPs procedure using a stainless-steel mesh was optimized. The optimal synthetic parameters were found to be one piece of stainless-steel mesh (22.5 cm2 in surface area) in 100 mL of a 1 mM precursor, precursor solution pH 4, and reaction temperature of 30°C. Under the optimal conditions, the as-synthesized (+)AuNPs were highly positively charged (+24.2 mV). Therefore, the as-synthesized (+)AuNPs act as a peroxidase mimic and provide a simple, fast, highly sensitive and selective colorimetric method for H2O2 detection with a detection limit of 0.06 mM in the linear range from 0.06 mM to 4.29 mM.