This paper presents a disposable biosensor to detect serum alcohol concentration. The proposed biosensor is fabricated using the cross-linking method to immobilize Alcohol Dehydrogenase (ADH) and Nicotinamide Adenine Dinucleotide (NAD(+)) on the screen-printed electrode modified with Meldola's Blue (MB) absorbed on Nafion. It is based on the electrocatalytic properties of MB as an electron transfer mediator, which can catalyze the oxidation of NADH to NAD(+) at a low oxidizing potential, thus avoiding interferences due to the presence of oxidizable substances in the real serum samples. The biosensor response for alcohol is investigated in terms of pH, buffer solution, temperature and some interferents. It presents the good specificity, reproducibility, stability, accuracy and provides a fast response. The biosensor has been satisfactorily used for the measurement of serum alcohol.