Background: Alzheimer’s disease (AD) is a common form of dementia, characterized by production and deposition of β-amyloid peptide in the brain. Thus, β-amyloid peptide is a potentially promising biomarker used to diagnose and monitor the progression of AD. Objective: The study aims to develop a biosensor based on a molecularly imprinted poly-pyrrole for detection of β-amyloid.Material and Methods: In this experimental study, an imprinted poly-pyrrole was employed as an artificial receptor synthesized by electro-polymerization of pyrrole on screen-printed carbon electrodes in the presence of β-amyloid. β-amyloid acts as a molecular template within the polymer. The biosensor was evaluated by cyclic voltammetry using ferro/ferricyanide marker. The parameters influencing the biosensor performance, including electro-polymerization cycle umbers and β-amyloid binding time were optimized to achieve the best biosensor sensitivity. Results: The β-amyloid binding affinity with the biosensor surface was evaluated by the Freundlich isotherm, and Freundlich constant and exponent were obtained as 0.22 ng mL-1 and 10.60, respectively. The biosensor demonstrated a detection limit of 1.2 pg mL-1. The biosensor was applied for β-amyloid determination in artificial cerebrospinal fluid. Conclusion: The biosensor is applicable for early Alzheimer’s disease detection.