Nonspecific interactions of conjugated polymers (CPs) with various proteins prove to be a major impediment for researchers when designing a suitable CP-based probe for the amplified and selective recognition of particular proteins in complex body fluids. Herein, a new strategy is presented for the precise and specific monitoring of clinically important serum albumin (SA) proteins at the nanomolar level using fluorescence resonance energy transfer (FRET)-modulated CP–surfactant ensembles as superior sensing materials. In brief, the newly designed color-tunable CP PF–DBT–Im undergoes intense aggregation with the surfactant sodium dodecyl sulfate (SDS), enabling drastic change in the emission color from violet to deep red due to intermolecular FRET. The emission of PF–DBT–Im/SDS ensembles then changed from deep red to magenta specifically on addition of SAs owing to the exclusive reverse FRET facilitated by synergistic effects of electrostatic interactions, hydrophobic forces, and the comparatively high intrinsic quantum yield of SAs. Interestingly, PF–DBT–Im itself could not differentiate SAs from other proteins, demonstrating the superiority of the PF–DBT–Im/SDS self-assembly over PF–DBT–Im. Finally, an affordable smartphone-integrated point-of-care (PoC) device is also fabricated as a proof-of-concept for the on-site and rapid monitoring of SAs, validating the potential of the system in long-term clinical applications.