The detection of microRNA plays an important role in early cancer diagnosis. Herein, a dual-mode electronic biosensor was developed for microRNA-21 (miRNA-21) detection based on gold nanoparticle-decorated MoS2 nanosheet (AuNPs@MoS2). A classical DNA “sandwich” structure was employed to construct MoS2-based electrochemical sensor, including capture DNA, target miRNA-21 and DNA-modified nanoprobe. Fe(CN)63-/4- and Ru(NH3)63+ were selected as electrochemical indicators to monitor the preparation process and evaluate the performance of MoS2-based electrochemical biosensor by electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV), respectively. Such MoS2-based biosensor exhibited excellent performance for miRNA-21 detection in the range from 10 fM to 1nM with detection limit of 0.78fM and 0.45fM for DPV and EIS technique, respectively. Furthermore, the proposed MoS2-based biosensor displayed high selectivity and stability, which could be used to determine miRNA-21 in human serum samples with satisfactory results. All data suggested that such MoS2-based nanocomposite may be a potential candidate for biosensing ranging from nucleic acid to protein detection. •A dual-mode electronic biosensor is developed for miRNA-21 detection based on AuNPs@MoS2.•DPV and EIS are used to detected miRNA-21 with high sensitivity and selectivity.•Suitable detection methods for nanomaterials-based biosensor could maximize the detection signal according to the nanomaterials' properties.