Dopamine (DA) is one of the most important catecholamine neurotransmitter molecules in the central nervous system (CNS). An abnormal level of DA in vivo can cause CNS diseases such as Parkinsonism and Schizophrenia. Thus, it is essential to develop an accurate and easy to use method for determining the level of DA in biological fluids such as urine and serum as a tool for clinical diagnostics as well as for pathological research. This work is the first ELISA application to detect DA in the presence of a high concentration of ascorbic acid (AA) and uric acid (UA) which are endogenous in urine. The LOD value of 1.26 x 10(-9) M and dynamic ranges of 3.16 x 10(-3) M to 3.16 x 10(-7) M were observed. It shows a good sensitivity with a broad dynamic range. Also, the competitive ELISA method for DA developed here showed no interference effect due to AA and UA, which are found in urine. The presence of AA and UA caused interference for DA determination by an electrochemical method because UA, AA, and DA have similar oxidation potential. Also, this selective and sensitive ELISA method can be made into an ELISA test kit which can be used to quantify the level of DA with ease and simplicity in clinics and laboratories.