Gliadins are allergenic proteins that can pose various risks to human health and are found in high amounts in the most commonly consumed foods. Gliadin, a component of gluten, triggers oxidative stress in celiac disease. Using sensitive analytical methods, especially in foods containing high amounts of these proteins, is crucial for food safety. Aptamer-based biosensors are widely preferred in electroanalytical methods. One of the promising approaches regarding biosensors is the studies in which aptamer-based sensors are combined with molecularly imprinted polymers. The antibody-like binding and ability of MIP to distinguish between molecules increases the method's selectivity. Using a platform modified with aptamer and molecularly imprinted polymer hybrid (MIP) as a new synthetic receptor film, this study presents a selective and sensitive label-free aptasensor for detecting a food allergy gliadin. For this purpose, screen printed gold electrode was used. The electrode surface was electrochemically coated with gold nanoparticles to bind the aptamer to the surface; then the aptamer-gliadin complex was dripped onto the surface and adhered to it, then, o-phenylene diamine monomer was used to synthesize a MIP surface. Gliadin was determined by using differential pulse voltammetry (DPV) in a wide range from 0.25 fg/mL to 1000 pg/mL. The developed method calculated LOD for gliadin determination as 0.011 fg/mL and LOQ as 0.034 fg/mL. The devised aptasensor was not only capable to the discrimination of the commonly found allergen compounds such as bovine serum albumin (BSA), casein, and ara-H1, but also it could detect the gliadin in spiked real samples such as gluten-free bread, crackers, cookies and brown rice cakes samples in the high recovery range of 98.1-104.6 %. The method could be a promising candidate for the sensitive determination of several allergens in food sample analysis.