Nanochannel-based plasmon-enhanced Raman scattering (PERS) substrates can simulate biological environments, revealing the recognition and conformation information on biomolecules in confined spaces. In this work, a metamaterial nanochannel-based PERS platform was constructed for highly sensitive analysis of DNA recognition to Hg2+ with the lowest Hg2+ concentration down to 1.0 pM. The established platform enables in situ monitoring of the thermodynamics and kinetics of DNA-Hg2+ recognition reaction in a confined nanospace. The recognition reaction in a nanospace shows good reversibility and specificity, and the isotherm follows well the Freundlich adsorption model. Compared to its folding on a rough Au nanofilm, the folding time of ssDNA-Rox decorated in nanochannels is remarkably increased, and the folding process can be tuned through varying the pore size and ionic strength. The presented PERS platform is promising for studying biomolecule-ion binding events and biomolecule conformation change under nanochannel-confined conditions.