We herein report a highly sensitive electrochemical sensor for glucose detection using carbon nanotubes grown in-situ at low temperatures on photolithographically defined gold microelectrode arrays printed on glass substrate (CNTs/Au MEA). One of the main advantages of the present design is its potential to monitor 64 samples individually for the detection of glucose. The selectivity of the fabricated MEA towards glucose detection is dramatically maintained via modification of CNTs/Au MEA by immobilizing glucose oxidase (GO_x) enzyme in the matrix of poly (paraphenylenediamine) (GO_x/poly (p-PDA)/CNTs/Au MEA). The electrocatalytic and electrochemical response of the proposed sensing platform towards glucose determination were examined via cyclic voltammetry and electrochemical impedance spectroscopy. The developed impedimetric biosensor exhibits a good linear response towards glucose detection i.e., 0.2-27.5 µM with sensitivity and detection limits of 168.03 kΩ^-1M^-1 and 0.2 ± 0.0014 μM, respectively. The proposed glucose biosensor shows excellent reproducibility, good anti-interference property and was successfully tested in blood serum samples. Further, applicability of the proposed sensor was successfully validated through HPLC. These results supported the viability of using such devices for the simultaneous detection of multiple electroactive biomolecules of physiological relevance.