Background and aims: Recently, in addition to numerous molecular methods available for diagnosis, methods based on the design of various biosensors considered. Some of these biosensors are Lateral flow tests based on gold nanoparticles. Considering the importance of the application of gold nanoparticles as markers in biosensors and also the need for the use of sugars in the construction of biosensors, this study was aimed to investigate the effects of various sugars (monosaccharides and disaccharides) on gold nanoparticles for the development of diagnostic biosensors. Methods: In this experimental study, after synthesis of gold nanoparticles chemically, by size, assessed maximum absorption at 520 nm, and concentration of nanoparticles by spectrophotometer and DLS method. Then, considering the importance of using sugars in the manufacture of biosensors, a concentration gradient of 5 to 500 mM glucose, fructose, sucrose, and trehalose, and their absorption spectra, after exposure to gold nanoparticles at 450 to 700 nm was detected. Results: The results of the spectrophotometric spectrum of synthesized nanoparticles in the presence and absence of sugars of monosaccharide and disaccharide indicate a decrease in the maximum absorption of nanoparticles after the application of a steady slope of sugar types on them and also concentration gradient of glucose, sucrose and trehalose sugars had no effect on the accumulation and aggregation of gold nanoparticles, while fructose at concentrations up to 50 mM leads to the accumulation of gold nanoparticles. Conclusion: Due to the necessity of using sugars in the production of bionanosensors, it seems that the use of ketone sugars, such as fructose, at concentrations of 50 mM upwards is not suitable for making biosensors.