A DNA tetrahedral scaffold-based platform for the construction of ECL biosensor is designed. DNA tetrahedron provides necessary space for TSP extending and subsequent assembly of luminol labelled Au NPs. Telomerase from HeLa cells was sensitively detected by the ECL enhancement from both CdS QDs and luminol. This work suggests that DNA tetrahedral scaffolds serve as an excellent choice for the construction of SPR-ECL system.