Platinum(Pt)/nanofibrous polyaniline(PANI) electrode was prepared by pulse galvanostatic method and characterized by scanning electron microscopy. The electrochemical behavior of L-cysteine at the Pt/nanofibrous PANI electrode was investigated by cyclic voltammetry. The results indicate that the pH value of the solution and the Pt loading of the electrode have great effect on the electrocatalytic property of the Pt/nanofibrous PANI electrode; the suitable Pt loading of the electrode is 600 μg/cm2 and the suitable pH value of the solution is 4.5 for investigating L-cysteine oxidation. The L-cysteine sensor based on the Pt/nanofibrous PANI electrode has a good selectivity, reproducibility and stability. The Pt/nanofibrous PANI electrode is highly sensitive to L-cysteine, and the linear calibration curve for the oxidation of L-cysteine can be observed in the range of 0.2-5.0 mmol/L.