Optimization of aptamer-triggered hybridization chain reaction for rapid visual ATP detection using gold nanoparticles

), which also support the functionality of the aptamer. This optimization allowed for a degree of HCR progression comparable to the traditional 24-hour incubation to be achieved within just 60 minutes. Subsequently, recognizing the inherent trade-off between the high ssDNA concentration required for fast HCR and the AuNP dispersion stability, we optimized the mixing ratio between the HCR product and the AuNP solution. Under these optimized conditions, the assay demonstrated the capability for rapid visual detection of ATP at the 100 µM level within a total assay time of 15 minutes, representing a significant acceleration compared to previously reported methods. Further improvements in sensitivity are anticipated through future fine-tuning of AuNP parameters and the use of post-reaction salt aggregation enhancers.