Hydrogel based substrate controllable release for lateral flow assay with automatic signal amplification

The limited sensitivity of conventional lateral flow assay (LFA) restricts its application in detecting trace targets. Although nanozyme-based LFAs could enhance detection performance significantly, they typically introduce additional steps, including the mixing and addition of the substrate for incubation, complicating the assay procedure. To circumvents this, we developed a hydrogel-integrated LFA platform (HG-LFA) by incorporating a sodium alginate (NaAlg) hydrogel network on a glass fiber membrane for encapsulating the chromogenic substrate 3,3'-diaminobenzidine (DAB). This three-dimensional network enables controllable release of DAB, automatically enhancing signal amplification with low background interference. Using SARS-CoV-2 E nucleic acid as a model target, the assay achieved a LOD of 0.146 nM, representing a 25-fold improvement over conventional Au@PtNPs-based LFA without signal amplification. After DAB catalysis, the signal enhancement of the HG-LFA that automatically releases the chromogenic substrate was comparable to that achieved by manual addition. In addition, the HG-LFA platform demonstrated high accuracy and stability in the detection of the target compound in biological matrix, offering a facile strategy well-suited for point-of-care testing (POCT) with high performance.