Highly sensitive label-free fluorescent detection of Hg2+ ions by DNA molecular machine-based Ag nanoclusters

Abstract

We present here a highly selective and sensitive label-free method to detect Hg²⁺ ions in aqueous solution by using DNA molecular machine-based fluorescent Ag nanoclusters (AgNCs). This mechanism is based on the Hg²⁺ ions triggering machine-like operations of DNA and the “product” of the machine being used to stabilize fluorescent AgNCs. In this method, a tailored DNA, containing a sequence for Hg²⁺ ions recognition, a sequence-specific nicking site for Nb BbvC I and a sequence complementary to the DNA as a template for the synthesis of fluorescent AgNCs, was firstly designed. In the presence of Hg²⁺ ions, the machine’s function operations were triggered. A series of machine-like operations, including replication, scission, and displacement then occurred with the addition of polymerase/dNTPs/Nb BbvC I, which manufactured lots of “product” DNA. The “product” DNA could act as a template for the preparation of fluorescent AgNCs. Thus the fluorescence of the AgNCs could be used as a signal transduction of this DNA machine, which was related to the concentration of the Hg²⁺ ions. The repeated synthesis of the “product” and its template effect for AgNCs synthesis led to signal amplification in the assay of Hg²⁺ ions. A linear response to the concentration of Hg²⁺ ions was observed in the range from 0.08 nM to 20 nM and a detection limit of 0.08 nM was obtained. By contrast, the operation of the machine could not be executed in an Hg²⁺ ion-free system. Moreover, the detection was not only label-free but also specific for Hg²⁺ ions without being affected by other metal ions.