Here, we proposed an enzyme-linked oligonucleotide assay (ELONA) to quantitatively detect periostin using periostin-specific two ssDNA aptamers in a sandwich manner. Patients with T H 2-high asthma have higher periostin levels in their blood. The two aptamers (PL2 trim and PL5 trim ) developed for the first time in this study showed specific binding affinity (PL2 trim K d = 10.76 nM, PL5 trim K d =36.97 nM) and a dual binding property that did not interfere with each other for periostin. Indeed, the sandwich ELONA using the dual binding property showed a meaningful detection of periostin, and showed the best detection performance when the PL2 trim and PL5 trim were used as capture and detection aptamer, respectively. In addition, the sensitivity of the sandwich assay was significantly increased by introducing streptavidin-poly horseradish peroxidase (SA-poly HRP). The established assay system was able to detect periostin within 3 hours and achieved low detection limits of 0.32 nM and 5.3 nM in buffer and serum spike conditions, respectively. Moreover, the aptasensor detected only periostin by distinguishing it from other blood proteins in buffer. Therefore, the sandwich ELONA could provide cost-effectiveness, high sensitivity, and superior specificity for periostin, which proves a possible use in discriminating T H 2-high asthmatics with high periostin levels.
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