Elevated serum high-sensitivity C-reactive protein (hs-CRP) and lipoprotein(a) [Lp(a)] levels are associated with the development of native coronary atherosclerosis. We aimed to establish a new method for the simultaneous detection of hs-CRP and Lp(a) to predict the development of atherosclerosis. A one-step time-resolved fluorescence immunoassay (TRFIA) with europium(III) (Eu3+) or samarium(III) (Sm3+) labels was established, and the performance of this TRFIA (in terms of sensitivity, specificity, accuracy and cutoff values) was evaluated using clinical serum samples and compared with those of registered kits. The sensitivity was 0.052 μg/mL for hs-CRP and 0.64 μg/mL for Lp(a). The intra-assay and inter-assay CVs were very low, ranging from 2.05% to 4.67% for hs-CRP and 2.42% to 6.43% for Lp(a). The cross-reactivities were very low (<0.34% and <2.65%, respectively) with five interferents. Additionally, there was a high Pearson coefficient between the present TRFIA method and the registered kits (R2 = 0.9967 and R2 = 0.9906, respectively). These data indicate that this study developed a TRFIA method that can be used for the quantitative detection of hs-CRP and Lp(a) in serum with high sensitivity, specificity and accuracy. This TRFIA provides a new method for predicting the development of atherosclerosis.

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