Entropy-driven multicolor DNA nanoflowers for simultaneous and rapid detection of multiple heavy metal ions in water

Heavy metal ions (HMIs), including Pb2+, Cu2+, and Hg2+, has shown great threat to the environment and public health worldwide owing to their high toxic at trace levels. However, conventional laboratory-based methods for analyzing HMIs require expensive instrumentation and complex operations limit their further application. In this study, we present a novel "two-step" approach based on entropy-driven multicolor DNA nanoflowers (DNFL) for the simultaneous ultrasensitive detection of three typical HMIs (Pb2+, Cu2+, and Hg2+). Through specific cleavage/competition reactions initiated by the target metal ions, a large number of single-stranded DNA (ssDNA) are produced, triggering the blooming of DNFL. Subsequently, with the assistance of fuel strands, an entropy-driven strand displacement reaction (ESDR) will further accelerate the conformational change of DNFL within one hour. Different fluorophores (FAM, ROX, and Cy5) attached to the opened DNFL serve as indicator molecules for the respective HMIs. After reasonable design and optimization, the developed fluorescence analysis method demonstrates excellent analytical performance (with limit of detection, 0.13 nM, 3.5 pM, 4.6 pM for Pb2+, Cu2+, and Hg2+, respectively) and can be applied to the simultaneous and rapid detection of multiple HMIs in water environment.