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Lateral Flow Assays (LFAs) are cost‐effective and widely utilized for rapid diagnostics, yet they often suffer from limited sensitivity. This study introduces a straightforward yet highly effective method to enhance LFAs performance by integrating a poly(ethylene glycol) diacrylate (PEGDA) hydrogel to create a "reaction trap." This hydrogel reaction trap optimizes the flow rate and reaction time at the sensing components, substantially improving assay performance. By applying various hydrogel concentrations (6%, 9%, 12%, 15%, and 18% w/v), significant enhancements across a range of detection systems are achieved. An optimized 18% hydrogel concentration shows a 1.5 times increase in sensitivity in the tested commercial pregnancy test. Additionally, 12% hydrogel concentration is tested in pregnancy, ovulation, and Coronavirus disease 2019 (COVID‐19) commercial kits, and the improvement reached up to a sevenfold increase in signal intensity. The enhancement in detection illustrates the profound impact of this simple modification and shows the importance of hydrogel concentration optimization to maximize detection improvement. These results demonstrate that hydrogel‐coated LFAs offer a scalable and highly effective solution for boosting the reliability and sensitivity of rapid diagnostics across different healthcare settings, with broad potential for global health diagnostics applications. [ABSTRACT FROM AUTHOR]

Hydrogen peroxide (H2O2) has been a fascinating target in various chemical, biological, clinical, and industrial fields. Several types of fluorescent protein-stabilized gold nanoclusters (protein-AuNCs) have been developed for sensitive and easy detection of H2O2. However, its low sensitivity makes is difficult to measure negligible concentrations of H2O2. Therefore, to overcome this limitation, we developed a horseradish peroxidase-encapsulated fluorescent bio-nanoparticle (HEFBNP), comprising bovine serum albumin-stabilized gold nanoclusters (BSA-AuNCs) and horseradish peroxidase-stabilized gold nanoclusters (HRP-AuNCs). The fabricated HEFBNP can sensitively detect H2O2 owing to its two properties. The first is that HEFBNPs have a continuous two-step fluorescence quenching mechanism, which comes from the heterogenous fluorescence quenching mechanism of HRP-AuNCs and BSA-AuNCs. Second, the proximity of two protein-AuNCs in a single HEFBNP allows a reaction intermediate (•OH) to rapidly reach the adjacent protein-AuNCs. As a result, HEFBNP can improve the overall reaction event and decrease the loss of intermediate in the solution. Due to the continuous quenching mechanism and effective reaction event, a HEFBNP-based sensing system can measure very low concentrations of H2O2 up to 0.5 nM and show good selectivity. Furthermore, we design a glass-based microfluidic device to make it easier use HEFBNP, which allowed us to detect H2O2 with the naked eye. Overall, the proposed H2O2 sensing system is expected to be an easy and highly sensitive on-site detection tool in chemistry, biology, clinics, and industry fields. [ABSTRACT FROM AUTHOR]