Juvenile idiopathic arthritis (JIA) is a wide group of diseases, characterized by synovial inflammation and joint tissue damage. Due to the delay in the implementation of biomarkers into clinical practice and the association with severe sequels, there is an imperative need for new JIA diagnosis strategies. Electrochemical biosensors based on screen-printed electrodes and peptides are promising alternatives for molecular diagnosis. In this work, a novel biosensor for detecting juvenile idiopathic arthritis (JIA) was developed based on the immobilization of the PRF+1 mimetic peptide, as recognition biological element, on the surface of screen-printed carbon electrode. This biosensor was able to discriminate the JIA positive and negative serum samples from different individuals using differential pulse voltammetry, presenting limits of detection and quantification in diluted samples of 1:784 (v/v) and 1:235 (v/v), respectively. Evaluation by electrochemical impedance spectroscopy showed RCT 3 times higher for JIA positive sample than for a pool of human serum samples from healthy individuals. Surface analysis of the biosensor by atomic force microscopy, after contact with JIA positive serum, presented great globular clusters irregularly distributed. The long-term stability of the biosensor was evaluated, remaining functional for over 40 days of storage (after storage at 8°C). Therefore, a simple, miniaturized and selective biosensor was developed, being the first one based on mimetic peptide and screen-printed carbon electrode, aiming at the diagnosis of the juvenile idiopathic arthritis in real serum samples.,Copyright © 2017 Elsevier B.V. All rights reserved.