<p>The convergence of nanotechnology and nucleic acid isothermal amplification (IA) is reshaping the molecular diagnostics landscape, bridging the gap between laboratory precision and field-scale testing. Unlike conventional PCR-based tests, AI techniques operate at constant temperature, allowing rapid, sensitive, and selective detection of nucleic acids with portable or even instrument-free instrumentation. Nanomaterials, including magnetic nanoparticles, gold nanostructures, and two-dimensional materials, among others, are now playing a critical role in enhancing AI systems. They facilitate efficient sample preparation through magnetic extraction, stabilize enzymatic reactions under suboptimal conditions, and serve as highly responsive transducers for generating optical or electrochemical signals. The integration of nanomaterials has also enabled advanced detection formats, from lateral flow assays and microfluidic cartridges to smartphone-coupled and CRISPR-assisted biosensors. These hybrid platforms combine high sensitivity and specificity with the simplicity required for point-of-care (POC) or resource-limited settings. Despite ongoing challenges in terms of standardization, reproducibility, and large-scale production, the analytical advantages of nano-assisted IA, such as accelerated workflows, multiplexing capabilities, and quantitative portability, pose this convergence as a keystone for next-generation diagnostic devices. This review examines recent advances, analytical implications, and future prospects for synergies between nanomaterials and IA, highlighting their transformative potential for accessible, rapid, and equitable molecular testing.</p>