Calibration curves for hs-CRP detection; 0.001, 0.01, 0.1, 1, 10, and 100 mg/L standard CRP was diluted from CRP stock solution and reacted with optimal capture antibody (4 mg/L) to perform this calibration. A) The best fit for the calibration curve is y = 0.51× + 2.63 with r 2 = 0.991 in the one-step format. The black solid round is the fluorescence intensities of individual hs-CRP concentrations and the red straight line is the fitted curve. B) The best fit for the calibration curve is y = 0.49× + 2.82 with r2 = 0.994 in the two-step format. The black solid square represents the fluorescence intensities of individual hs-CRP concentrations with the red straight line representing the fitted curve. Error bars indicate the standard errors of 3 independent experiments.