A novel QD-based immunoassay on a paper-based lateral flow system has been developed to quantitatively detect C-reactive protein (CRP). Different standard CRP antigens from 1 to 200 μg mL-1 were diluted 200-fold and only 60 μL diluted sample were needed to load onto the sample pad. The QD fluorescence signals on the test line and the control line were able to be observed within 3 min after the initiation of assay, and the limit of detection was as sensitive as 0.30 ng mL-1 by measuring the fluorescence intensity immediately afterwards with fluorescence immunoassay analyzer. The linearity on the detection of QD fluorescence signals has been established well in the range of 0.5 ng mL-1 and 1 μg mL-1 for CRP. The precision of the assay has been confirmed for low coefficient of variation (CV), satisfying less than 15% (intra-assay and inter-assay), and the accuracy of assay meets the requirements with the mean recovery of the control was 102.63%. These results indicated that such newly developed platform was reliable with high sensitivity, rapidness, and could cover a broad range of target concentrations. Furthermore, a total of 135 human serum clinical samples with inflammation or infection with the concentration of CRP from 0.2 to 200 μg mL-1 has been used to check the performance of this QD-based LFIA, it correlated very well with Roche Tina-quant CRP (Latex) (r = 0.966, n = 135).