In this study, a metal enhanced fluorescence (MEF) approach using the homemade Ag nanowires for enzyme free and electrodeless detection of direct bilirubin (d-BLR) was successfully developed. The average diameter and length of the fabricated Ag nanowires were measured to be around 80 nm and 25 μm, respectively. Solvent effect experiments found that EtOH is more suitable as the solvent for d-BLR. The experimental results found that the linear detection range is from 1 to 10 μM, which can be used for detecting abnormal d-BLR concentration from 5 to 10 μM. The MEF approach not only enhanced the bilirubin fluorescence, but also improved the linearity of the standard curve. Furthermore, we also found that the fluorescence intensity of the chip-based MEF is much higher than that of the solution-based MEF. It is believed that the proposed enzyme free and electrodeless d-BLR detection platform is highly feasible for clinical applications.