Enzyme-free colorimetric detection of MicroRNA-21 using metal chelator as label for signal generation and amplification.

MicroRNAs (miRNAs) were reported to be potential tumor markers for early diagnosis of cancer. Due to its short sequence, low expression level and high susceptibility to degradation, the stable and sensitive detection method of miRNAs is arduous to establish. In this work, we designed a metal chelator (ethylenediamine tetraacetic acid disodium salt, EDTA•2Na) labeled oligonucleotides as the plasmonic signal supraregulator probe to control the generation of gold nanoparticles (AuNPs). Based on another complementary oligonucleotides of target miRNA labeling SiO2 microparticles (SiO2MPs) as the detecting platform, EDTA•2Na labeled oligonucleotide probes were immobilized on the SiO2 platform through the sandwich structure in the presence of target miRNA. The sandwich chelating device could further chelate Au3+ to regulate the generation of AuNPs, resulting in colorimetric signal to qualitatively and quantitatively detect the concentration of microRNA-21 (miR-21). The results indicate that the proposed metal chelator -labeled signal amplification method has outstanding sensitivity (LOD = 8.9 fM) and excellent stability, which will be benefit for the early accurate diagnosis of miRNAs.

[1]  D. Moon,et al.  Synthesis of LaNiO3 perovskite using an EDTA-cellulose method and comparison with the conventional Pechini method: application to steam CO2 reforming of methane , 2016 .

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

[3]  W. A. Norvell Reactions of Metal Chelates in Soils and Nutrient Solutions , 2018, Micronutrients in Agriculture.

[4]  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.

[5]  Ying Zhang,et al.  NIR‐Remote Selected Activation Gene Expression in Living Cells by Upconverting Microrods , 2016, Advanced materials.

[6]  R. Shiekhattar,et al.  The Microprocessor complex mediates the genesis of microRNAs , 2004, Nature.

[7]  Wei Wen,et al.  Enzyme catalytic amplification of miRNA-155 detection with graphene quantum dot-based electrochemical biosensor. , 2016, Biosensors & bioelectronics.

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

[9]  Wei Li,et al.  MicroRNA detection by microarray , 2009, Analytical and bioanalytical chemistry.

[10]  K. Czaplinski,et al.  Exportin 5 is a RanGTP-dependent dsRNA-binding protein that mediates nuclear export of pre-miRNAs. , 2004, RNA.

[11]  Mark M. Davis,et al.  miR-181a Is an Intrinsic Modulator of T Cell Sensitivity and Selection , 2007, Cell.

[12]  S. Abolmaali,et al.  miR-21, An Oncogenic Target miRNA for Cancer Therapy: Molecular Mechanisms and Recent Advancements in Chemo and Radio-resistance. , 2017, Current gene therapy.

[13]  C. Perou,et al.  A custom microarray platform for analysis of microRNA gene expression , 2004, Nature Methods.

[14]  É. Várallyay,et al.  MicroRNA detection by northern blotting using locked nucleic acid probes , 2008, Nature Protocols.

[15]  S. McGrath,et al.  Leaching of heavy metals from contaminated soils using EDTA. , 2001, Environmental pollution.

[16]  F. Tang,et al.  Maternal microRNAs are essential for mouse zygotic development. , 2007, Genes & development.

[17]  H. Lodish,et al.  Micromanagement of the immune system by microRNAs , 2008, Nature Reviews Immunology.

[18]  Chunhai Fan,et al.  A dumbbell probe-mediated rolling circle amplification strategy for highly sensitive microRNA detection , 2010, Nucleic acids research.

[19]  Yuriy Gusev,et al.  Real-time expression profiling of microRNA precursors in human cancer cell lines , 2005, Nucleic acids research.

[20]  N. Sandhyarani,et al.  A novel electrochemical sensor surface for the detection of hydrogen peroxide using cyclic bisureas/gold nanoparticle composite. , 2011, Biosensors & bioelectronics.

[21]  Chunhai Fan,et al.  Visual cocaine detection with gold nanoparticles and rationally engineered aptamer structures. , 2008, Small.

[22]  R. Stephens,et al.  Unique microRNA molecular profiles in lung cancer diagnosis and prognosis. , 2006, Cancer cell.

[23]  Zong Dai,et al.  Asymmetric exponential amplification reaction on a toehold/biotin featured template: an ultrasensitive and specific strategy for isothermal microRNAs analysis , 2016, Nucleic acids research.

[24]  H. Blöcker,et al.  Predicting DNA duplex stability from the base sequence. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Gurman Singh Pall,et al.  Carbodiimide-mediated cross-linking of RNA to nylon membranes improves the detection of siRNA, miRNA and piRNA by northern blot , 2007, Nucleic acids research.

[26]  Vladimir Benes,et al.  A sensitive array for microRNA expression profiling (miChip) based on locked nucleic acids (LNA). , 2006, RNA.

[27]  D. Bartel MicroRNAs: Target Recognition and Regulatory Functions , 2009, Cell.

[28]  M. Grzelczak,et al.  Using gold nanoparticles to detect single-nucleotide polymorphisms: toward liquid biopsy , 2020, Beilstein journal of nanotechnology.

[29]  R. Álvarez-Puebla,et al.  Cancer Diagnosis through SERS and Other Related Techniques , 2020, International journal of molecular sciences.

[30]  Lehui Lu,et al.  Hydrogen-bonding recognition-induced color change of gold nanoparticles for visual detection of melamine in raw milk and infant formula. , 2009, Journal of the American Chemical Society.

[31]  Yun Zhang,et al.  Study on adsorption mechanism of Pb(II) and Cu(II) in aqueous solution using PS-EDTA resin , 2010 .

[32]  Kai Zhang,et al.  Sensitive detection of microRNA in complex biological samples by using two stages DSN-assisted target recycling signal amplification method. , 2017, Biosensors & bioelectronics.

[33]  D. A. Fleming,et al.  Size-controlled synthesis of gold nanoparticles via high-temperature reduction. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[34]  K. Livak,et al.  Real-time quantification of microRNAs by stem–loop RT–PCR , 2005, Nucleic acids research.

[35]  S. Rai,et al.  Plasma MiR-21: A Potential Diagnostic Marker of Colorectal Cancer , 2012, Annals of surgery.

[36]  L. Chen,et al.  A visual detection of hydrogen peroxide on the basis of Fenton reaction with gold nanoparticles. , 2010, Analytica chimica acta.

[37]  Nóra Varga,et al.  Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes. , 2004, Nucleic acids research.