Due to its unique programmability, nanosized structure, and biocompatible properties, deoxyribonucleic acid (DNA) has attracted increasing attention for the construction of versatile nanostructures and nanomaterials. This review summarizes the recent developments in DNA-templated fluorescent nanomaterials, including DNA-stabilized quantum dots (QDs) and DNA-templated metal nanoclusters (NCs), as well as their applications in bioanalysis. These fluorescent nanomaterials not only have good fluorescence properties but also exhibit excellent performance in DNA recognition, which greatly expands the range of their bioanalytical applications. Finally, we discuss some current challenges and future work in this field, with the goal of further promoting the potential applications and developments of DNA-templated fluorescent nanomaterials in biochemical analysis.