In the brain and central nervous system, dopamine plays a crucial role as a neurotransmitter or a local chemical messenger for interneuronal communication. Dopamine is associated with renal, hormonal, and cardiovascular systems. Additionally, dopamine dysfunction is known to cause serious illnesses, such as Parkinson's disease and Alzheimer's disease. Therefore, dopamine detection is essential for medical diagnosis and disease prevention and requires a novel strategy with high sensitivity and selectivity and a rapid response. Herein, we present a novel human dopamine receptor (hDRD1)-conjugated multidimensional conducting polymer nanofiber (NF) membrane for the selective and sensitive detection of dopamine. The membrane, which consists of multidimensional carboxylated poly(3,4-ethylenedioxythiophene) (MCPEDOT) NFs with nanorods, is used as a transistor in a liquid-ion gated field-effect transistor (FET)-based biosensor. Interestingly, hDRD1 is first expressed in Escherichia coli before it is immobilized onto the MCPEDOT NF. The hDRD1-MCPEDOT NF-based FET exhibits a rapid real-time response (<2 s) with high dopamine selectivity and sensitivity performance (approximately 100 fM). Furthermore, this FET device can be integrated into a poly(dimethylsiloxane)-based microfluidic system and also can retain its high performance in the integrated system, which results in the generation of large-scale dopamine biosensors with a novel geometry.