In this context a simple and rapid method for trace amounts of morphine detection has been described based on electrochemical methods of pencil graphite electrode (PGE) which was constructed by using Multi-wall carbon nanotubes (MWCNT), molecularly imprinted polymer (MIP) and gold-nanoparticles. In the electropolymerization process, selective MIP was produced using Methyltrimethoxysilane and Tetraethylorthosilicate as monomer and cross-linker respectively on modified electrode in NaOH solution. The structural morphology of the modified electrode was characterized by field emission scanning electron microscopy (FE-SEM) technique. Various types of electrochemical methods containing cyclic voltammetry (CV), square wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS) were employed to probe the characteristics of the constructed electrode toward morphine. After optimization of several effective parameters, the calibration curve was obtained by SWV which increased linearly dependent on the MO concentration over the range of 0.008 to 0.1 and 0.1 to 5 ?mol L -1 in two linear fragments vs. Ag/AgCl. The sensitivity for morphine detection was improved greatly and the detection limit was 2.85 nM. The relative standard deviation (RSD) for six replicate determinations of 0.05 ?mol L -1 morphine was found to be 5.46% (S/N = 6). Finally, the ability of the electrochemical sensor was successfully applied for determination of target in real samples such as human urine and plasma that was succeeded with satisfactory results. The results indicated that the proposed electrode exhibited high sensitivity and stability for detecting morphine.