Highly sensitive and selective strategy for microRNA detection based on WS2 nanosheet mediated fluorescence quenching and duplex-specific nuclease signal amplification.

MicroRNAs (miRNAs) play vital roles in physiologic and pathologic processes and are significant biomarkers for disease diagnostics and therapeutics. However, rapid, low-cost, sensitive, and selective detection of miRNAs remains a challenge because of their short length, sequence homology, and low abundance. Herein, we report for the first time that WS2 nanosheet can exhibit differential affinity toward short oligonucleotide fragment versus ssDNA probe and act as an efficient quencher for adsorbed fluorescent probes. This finding is utilized to develop a new strategy for simple, sensitive, and selective detection of miRNA by combining WS2 nanosheet based fluorescence quenching with duplex-specific nuclease signal amplification (DSNSA). This assay exhibits highly sensitive and selective with a detection limit of 300 fM and even discriminate single-base difference between the miRNA family members. The result indicates that this simple and cost-effective strategy holds great potential application in biomedical research and clinical diagnostics.

[1]  A. Radenović,et al.  Single-layer MoS2 transistors. , 2011, Nature nanotechnology.

[2]  Jian-hui Jiang,et al.  Graphene fluorescence resonance energy transfer aptasensor for the thrombin detection. , 2010, Analytical chemistry.

[3]  F. Slack,et al.  Oncomirs — microRNAs with a role in cancer , 2006, Nature Reviews Cancer.

[4]  Mrinmoy De,et al.  Ligand conjugation of chemically exfoliated MoS2. , 2013, Journal of the American Chemical Society.

[5]  A. Splendiani,et al.  Emerging photoluminescence in monolayer MoS2. , 2010, Nano letters.

[6]  C. Croce,et al.  MicroRNA gene expression deregulation in human breast cancer. , 2005, Cancer research.

[7]  Label-free detection of microRNA: two-step signal enhancement with a hairpin-probe-based graphene fluorescence switch and isothermal amplification. , 2013, Chemistry.

[8]  Chunhai Fan,et al.  Single-layer MoS2-based nanoprobes for homogeneous detection of biomolecules. , 2013, Journal of the American Chemical Society.

[9]  Qian Wang,et al.  High specific and ultrasensitive isothermal detection of microRNA by padlock probe-based exponential rolling circle amplification. , 2013, Analytical chemistry.

[10]  Xi Chen,et al.  Graphene oxide-protected DNA probes for multiplex microRNA analysis in complex biological samples based on a cyclic enzymatic amplification method. , 2012, Chemical communications.

[11]  H. Horvitz,et al.  MicroRNA expression profiles classify human cancers , 2005, Nature.

[12]  B. Fang,et al.  WS2 nanosheets as a highly efficient electrocatalyst for hydrogen evolution reaction , 2012 .

[13]  Jing Zhang,et al.  Highly sensitive multiple microRNA detection based on fluorescence quenching of graphene oxide and isothermal strand-displacement polymerase reaction. , 2012, Analytical chemistry.

[14]  Yu-Qiang Liu,et al.  One-step, multiplexed fluorescence detection of microRNAs based on duplex-specific nuclease signal amplification. , 2012, Journal of the American Chemical Society.

[15]  Huang-Hao Yang,et al.  A graphene platform for sensing biomolecules. , 2009, Angewandte Chemie.

[16]  Hua Zhang,et al.  Single-layer MoS2 phototransistors. , 2012, ACS nano.

[17]  Chun-yang Zhang,et al.  Sensitive detection of microRNA with isothermal amplification and a single-quantum-dot-based nanosensor. , 2012, Analytical chemistry.

[18]  Po-Jung Jimmy Huang,et al.  Molecular beacon lighting up on graphene oxide. , 2012, Analytical chemistry.

[19]  Yan Wang,et al.  Detection of microRNA by fluorescence amplification based on cation-exchange in nanocrystals. , 2009, Analytical chemistry.

[20]  B. Ye,et al.  Attomolar ultrasensitive microRNA detection by DNA-scaffolded silver-nanocluster probe based on isothermal amplification. , 2012, Analytical chemistry.

[21]  C. Croce,et al.  MicroRNA signatures in human cancers , 2006, Nature Reviews Cancer.

[22]  Mihaela Zavolan,et al.  Virus-encoded microRNAs: novel regulators of gene expression. , 2006, Trends in microbiology.

[23]  D. Bartel MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[24]  Hisato Yamaguchi,et al.  Enhanced catalytic activity in strained chemically exfoliated WS₂ nanosheets for hydrogen evolution. , 2012, Nature Materials.

[25]  Daniel B. Martin,et al.  Circulating microRNAs as stable blood-based markers for cancer detection , 2008, Proceedings of the National Academy of Sciences.

[26]  H. Wen,et al.  Control of Schottky barriers in single layer MoS2 transistors with ferromagnetic contacts. , 2013, Nano letters.

[27]  S. Lukyanov,et al.  A novel method for SNP detection using a new duplex-specific nuclease from crab hepatopancreas. , 2002, Genome research.