Facile construction of a highly sensitive DNA biosensor by in-situ assembly of electro-active tags on hairpin-structured probe fragment

An ultrasensitive DNA biosensor has been developed through in-situ labeling of electroactive melamine-Cu2+ complex (Mel-Cu2+) on the end of hairpin-like probe using gold nanoparticles (AuNPs) as the signal amplification platform. The 3′-thiolated hairpin-like probe was first immobilized to the gold electrode surface by the Au-S bond. The AuNPs were then tethered on the free 5′-end of the immobilized probe via the special affinity between Au and the modified -NH2. Followed by, the Mel and Cu2+ were assembled on the AuNPs surface through Au-N bond and Cu2+-N bond, respectively. Due to the surface area and electrocatalytic effects of the AuNPs, the loading amount and electron transfer kinetic of the Mel-Cu2+ were enhanced greatly, resulting in significantly enhanced electrochemical response of the developed biosensor. Compared with the synthesis process of conventional electroactive probe DNA accomplished by homogeneous method, the method presented in this work is more reagent- and time-saving. The proposed biosensor showed high selectivity, wide linear range and low detection limit. This novel strategy could also be extended to the other bioanalysis platforms such as immunosensors and aptasensors.

[1]  Yaping Hu,et al.  Universal surface-enhanced Raman scattering amplification detector for ultrasensitive detection of multiple target analytes. , 2014, Analytical chemistry.

[2]  Elena E Ferapontova,et al.  "Off-on" electrochemical hairpin-DNA-based genosensor for cancer diagnostics. , 2011, Analytical chemistry.

[3]  A. Bond,et al.  Highly selective and sensitive DNA assay based on electrocatalytic oxidation of ferrocene bearing zinc(II)-cyclen complexes with diethylamine. , 2010, Journal of the American Chemical Society.

[4]  Chun-Yang Zhang,et al.  Improved sensitivity for the electrochemical biosensor with an adjunct probe. , 2010, Analytical chemistry.

[5]  E. Ferapontova,et al.  Electrochemistry of weakly adsorbed species: Voltammetric analysis of electron transfer between gold electrodes and Ru hexaamine electrostatically interacting with DNA duplexes , 2014 .

[6]  Arne Thomas,et al.  Catalyst-free preparation of melamine-based microporous polymer networks through Schiff base chemistry. , 2009, Journal of the American Chemical Society.

[7]  J. Reboud,et al.  Rapid ultrasonic isothermal amplification of DNA with multiplexed melting analysis – applications in the clinical diagnosis of sexually transmitted diseases. , 2015, Chemical communications.

[8]  Jun‐Jie Zhu,et al.  Graphene quantum dots as fluorescence probes for turn-off sensing of melamine in the presence of Hg(2+). , 2014, ACS applied materials & interfaces.

[9]  D. Bélanger,et al.  Covalent grafting of aminated compounds on Vulcan XC72R by melamine in situ diazotization , 2012 .

[10]  J. Chalker,et al.  Melamine and melamine-formaldehyde polymers as ligands for palladium and application to Suzuki-Miyaura cross-coupling reactions in sustainable solvents. , 2014, The Journal of organic chemistry.

[11]  E. Laviron.  General expression of the linear potential sweep voltammogram in the case of diffusionless electrochemical systems , 1979 .

[12]  John H T Luong,et al.  Hairpin DNA as a biobarcode modified on gold nanoparticles for electrochemical DNA detection. , 2015, Analytical chemistry.

[13]  Shaorong Liu,et al.  Charging YOYO-1 on Capillary Wall for Online DNA Intercalation and Integrating This Approach with Multiplex PCR and Bare Narrow Capillary–Hydrodynamic Chromatography for Online DNA Analysis , 2015, Analytical chemistry.

[14]  A. Ellington,et al.  Reagentless, Ratiometric Electrochemical DNA Sensors with Improved Robustness and Reproducibility , 2014, Analytical chemistry.

[15]  F. Gao,et al.  Development of a novel electrochemical DNA biosensor based on elongated hexagonal-pyramid CdS and poly-isonicotinic acid composite film. , 2014, Biosensors & bioelectronics.

[16]  Zhiwei Zhu,et al.  Electrochemical sensor for melamine based on its copper complex. , 2010, Chemical communications.

[17]  Xiaolei Zuo,et al.  Rational designed bipolar, conjugated polymer-DNA composite beacon for the sensitive detection of proteins and ions. , 2015, Analytical chemistry.

[18]  V. Chiș,et al.  FTIR, FT-Raman, SERS and DFT study on melamine , 2012 .

[19]  Jian Ji,et al.  Rapid and sensitive detection of foodborne pathogenic bacteria (Staphylococcus aureus) using an electrochemical DNA genomic biosensor and its application in fresh beef. , 2014, Journal of agricultural and food chemistry.

[20]  Agostina Congiu Castellano,et al.  New marker of tumor cell death revealed by ATR-FTIR spectroscopy , 2011, Analytical and bioanalytical chemistry.

[21]  Erin K. Grey,et al.  Rapid molecular detection of invasive species in ballast and harbor water by integrating environmental DNA and light transmission spectroscopy. , 2015, Environmental science & technology.

[22]  Xiaojun Zhang,et al.  A simple, fast, and sensitive assay for the detection of DNA, thrombin, and adenosine triphosphate based on Dual-Hairpin DNA structure. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[23]  Pierre-Louis Taberna,et al.  Electrochemical quartz crystal microbalance (EQCM) study of ion dynamics in nanoporous carbons. , 2014, Journal of the American Chemical Society.

[24]  S. P. Samijlenko,et al.  ATR-IR spectroscopy as applied to nucleic acid films. , 2005, Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy.

[25]  Zhenyu Lin,et al.  Ultraselective homogeneous electrochemical biosensor for DNA species related to oral cancer based on nicking endonuclease assisted target recycling amplification. , 2015, Analytical chemistry.

[26]  J. Roseiro,et al.  Selecting low-cost carbon sources for carotenoid and lipid production by the pink yeast Rhodosporidium toruloides NCYC 921 using flow cytometry. , 2014, Bioresource technology.

[27]  C. Chaix,et al.  A thermodynamic study of ferrocene modified hairpin oligonucleotides upon duplex formation: applications to the electrochemical detection of DNA , 2009 .

[28]  L. Alvarez-Cohen,et al.  Development of a fluorescence-activated cell sorting method coupled with whole genome amplification to analyze minority and trace Dehalococcoides genomes in microbial communities. , 2015, Environmental science & technology.

[29]  Catherine J. Murphy,et al.  Seed‐Mediated Growth Approach for Shape‐Controlled Synthesis of Spheroidal and Rod‐like Gold Nanoparticles Using a Surfactant Template , 2001 .

[30]  Ulrich Janus,et al.  In-situ atomic force microscopy (AFM) imaging: influence of AFM probe geometry on diffusion to microscopic surfaces. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[31]  Arica A Lubin,et al.  Single-step electronic detection of femtomolar DNA by target-induced strand displacement in an electrode-bound duplex , 2006, Proceedings of the National Academy of Sciences.

[32]  J. Anzai,et al.  Use of polymeric indicator for electrochemical DNA sensors: poly(4-vinylpyridine) derivative bearing [Os(5,6-dimethyl-1,10-phenanthroline)2Cl]2+. , 2004, Analytical chemistry.

[33]  H. Korri-Youssoufi,et al.  E-DNA sensor of Mycobacterium tuberculosis based on electrochemical assembly of nanomaterials (MWCNTs/PPy/PAMAM). , 2015, Analytical chemistry.

[34]  Chunhai Fan,et al.  Electrochemical interrogation of conformational changes as a reagentless method for the sequence-specific detection of DNA , 2003, Proceedings of the National Academy of Sciences of the United States of America.