HIV biosensors for early diagnosis of infection: The intertwine of nanotechnology with sensing strategies.

Silver nanoclusters stabilized with DNA scaffolds, with excellent physical and chemical properties, have emerged as a versatile tool in biomedical systems. Silver nanoclusters as a late-model nanomaterial with a small size less than 2 nm shows good stability and strong fluorescence. By using DNA with diverse sequence and structures, soluble DNA-templated silver nanoclusters are facilely prepared. They possess tunable fluorescence emission and are suitable with multifunctional designs. Extensive efforts have been put into the application potential of this biocompatible material for biosensing, bioimaging and therapy. Here we are committed to outline the recent advances of DNA-templated silver nanoclusters in biomedical application. The future prospect and challenges are then discussed considering rising progress. Though there is still a long way for illuminating the mechanism behind traits and conducting further clinical trials, we believe DNA-templated silver nanoclusters’ unprecedented unique properties will finally benefit medical progress.

Recent advances in the bioanalytical and biomedical applications of DNA-templated silver nanoclusters

Metal quantum clusters are evolving as excellent systems for a wide range of biological applications due to their small size (∼2 nm), tunable optical properties, including optical absorption, photoluminescence (UV to NIR), nonlinear optical properties (two-photon absorption, two-photon fluorescence, and second/third harmonic generation), ultrafast dynamics (relaxation kinetics, electron–phonon coupling, and radiative emission), and magnetism. These excellent properties have resulted in their use in a broad range of applications, including the sensing of ions (heavy metal ions, anions), biomolecules (proteins, DNA, miRNA, and enzymes), biological cells, diagnosis, and therapy. This article presents an introduction to metal quantum clusters, including a brief history of research in this system and an overview of the existing theories to understand their properties. We also discuss the synthesis methods, the various properties of quantum clusters and present a broad and updated overview of the applications of metal quantum clusters in biology.

Fluorescent metal quantum clusters: an updated overview of the synthesis, properties, and biological applications

miRNAs have attracted increased attention as biomarkers, which have strong associations with diseases like cancers. Herein, we have constructed DNA four-way junction architecture on the surface of electrode for ultrasensitive electrochemical detection of miRNA. Isothermal duplex-specific nuclease catalyzed reaction and nicking endonuclease catalyzed reaction are performed on the synthesized novel magnetic nanomaterials and the formed DNA junctions, respectively. Besides the enzymatic reaction cycles, TCEP mediated electrochemical amplification is integrated for improvement of the sensitivity. This method not only achieves attomolar sensitivity, high selectivity and practical utility have also been confirmed. Therefore, the proposed approach is supposed to provide a powerful tool for miRNA related biological researches and clinical applications.

Manipulations of DNA four-way junction architecture and DNA modified Fe3O4@Au nanomaterials for the detection of miRNA

Sensitive detection of cancer biomarker is of great importance for early accurate diagnostics and therapy of cancer patients. Herein, a label-free and enzyme-free ratiometric fluorescence biosensor is developed for the detection of carcinoembryonic antigen (CEA) based on a recycling amplification strategy. CEA could trigger the release of G-quadruplex sequences from a DNA machine in the presence of hairpin DNA (H-DNA) with an aptamer fragment and the split of the silver nanoclusters (AgNCs) pair in the presence of fuel DNA (F-DNA), resulting in an enhanced fluorescence of Thioflavin T (THT) and a decline fluorescence of the AgNCs pair. The novel ratiometric fluorescence biosensor prevents the cumbersome functionalization process and the shortcoming of enzyme-based amplification strategy. Under the optimal conditions, the present biosensor achieves a detection limit of 0.1 ng mL−1 and a linear range of 0.5–20 ng mL−1 for CEA, which is superior to the single-signal strategy based on THT or AgNCs. The practical applicability of the present biosensor is demonstrated by the detection of CEA in human serum with satisfactory results, thus it shows great potential in future clinical diagnosis.

A label-free and enzyme-free ratiometric fluorescence biosensor for sensitive detection of carcinoembryonic antigen based on target-aptamer complex recycling amplification

A label-free and sensitive fluorescence biosensing platform for human immunodeficiency virus gene (HIV-DNA) detection has been fabricated based on luminescent DNA-scaffolded silver nanoclusters (DNA/AgNCs) and autonomous exonuclease III (Exo III)-assisted recycling signal amplification. One long-chain DNA (X-DNA) molecule can hybridize with two assistant DNA (F-DNA) molecules and one HIV-DNA molecule; after Exo III digests X-DNA to liberate F-DNA and HIV-DNA. F-DNA combines with P-DNA (template of DNA/AgNCs), accordingly, P-DNA is cut and the fluorescence of the system is quenched. This assay can finish in one-step without any labelling of the DNA chain or complex construction, and the strategy is sensitive with the detection limit as low as 35 pM. At the same time, the approach exhibits good selectivity even against a single base mismatch. What's more, the method is able to monitor HIV-DNA in real human serum samples; it holds great potential for early diagnosis in gene-related diseases.

A new label-free fluorescent sensor for human immunodeficiency virus detection based on exonuclease III-assisted quadratic recycling amplification and DNA-scaffolded silver nanoclusters

A facile, highly selective biosensor was used for ATP quantitative analysis. Here we introduce two hairpin probes as the matrix. While Exo III digested their duplexes, the blocked G-quadruplex released. Then N-methyl mesoporphyrin IX (NMM) integrated with G-quadruplex to produce strong fluorescence. But when ATP was added, it triggered the T4 DNA ligase to link the two hairpins. As a result, Exo III could not hydrolyze the hairpins leading to a low signal out. With this method, the detection limit reached 3.75 nM, and linear range 5–300 nM for ATP. This method has good selectivity and specificity without a tedious treatment and label process.

Free-labelled fluorescent method for ATP detection assisted by T4 DNA ligase

Detection of microRNA using enzyme-assisted amplifying and DNA-templated silver nanoclusters signal-off fluorescence bioassay.

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A new label-free fluorescent sensor for human immunodeficiency virus detection based on exonuclease III-assisted quadratic recycling amplification and DNA-scaffolded silver nanoclusters

Free-labelled fluorescent method for ATP detection assisted by T4 DNA ligase

Detection of microRNA using enzyme-assisted amplifying and DNA-templated silver nanoclusters signal-off fluorescence bioassay.