High-Sensitivity DNA Aptasensors for Detecting Salivary Biomarker S100A7 in Heart Failure.

Early detection of heart failure (HF) is vital for improving patient outcomes, lowering hospital readmission rates, and enabling prompt treatment. We present the first high-affinity DNA aptamer for the salivary HF biomarker S100A7 and its application in highly sensitive, noninvasive diagnostic tests. Iterative truncation of the initial 82-nt aptamer (17-82) produced a 43-nt core (17-43) with a binding affinity of 27 nM, which was further enhanced to 5.5 nM through dimerization. Biochemical and mutational studies confirmed that 17-43 adopts a G-quadruplex structure, which is essential for S100A7 recognition and resistance to enzymatic degradation in human saliva. Incorporating 17-43 into sandwich aptamer-ELISA and hybrid aptamer-antibody ELISA assays allowed detection of recombinant S100A7 in human saliva with limits of detection (LOD) of 7.4 ng mL-1 (0.6 nM) and 29 pg/mL (2.2 pM), respectivelyoutperforming commercial immunoassays in both sensitivity and dynamic range. The hybrid assay maintained its full performance after 2.5 months of room temperature storage. Additionally, a biolayer interferometry (BLI) sensor with 17-43 quantified S100A7 in patient saliva (n = 3), achieving a LOD of 3.2 ng mL-1 (0.3 nM) with a total assay time of less than 20 min. The aptamer's stability, high specificity, and versatility across biosensing platforms establish it as a promising tool for noninvasive heart failure diagnostics, laying the groundwork for portable aptamer-based biosensors for multiplexed monitoring of HF biomarkers.