In this report, we present a new detection method for blood glucose, using gold nanorod SERS, a surface enhanced Raman scattering probe embedded in two component self-assembled monolayers (SAMs). Gold nanorod particles and a gold coated slide surface were modified with the two component SAMs consisting of 3-mercaptophenylboronic acid (3-MBA) and 1-decanethiol (1-DT). The immobilization of 3-MBA/1-DT surface-functionalized gold nanoparticles onto 3-MBA/1-DT modified gold-coated slide surfaces was achieved by the cooperation of hydrophobic forces. Two component SAM functionalized substrates were used as SERS probes, by means of the boronic acid and the alkyl spacer functional groups that serve as the molecular recognition and penetration agents, respectively. The SERS platform surface was characterized by cyclic voltammetry, contact angle measurements, AFM (atomic force microscopy) and Raman spectroscopy. Optimum values of the parameters such as pH, time and (3-MBA/1-DT) molar ratio were also examined for the glucose determination. The analytical performance was evaluated and linear calibration graphs were obtained in the glucose concentration range of 2-16 mM, which is also in the range of the blood glucose levels, and the detection limit was found to be 0.5 mM. As a result, the SERS platform was also used for the determination of glucose in plasma samples.