An ultrasensitive photoelectrochemical nucleic acid biosensor

A simple and ultrasensitive procedure for non-labeling detection of nucleic acids is described in this study. It is based on the photoelectrochemical detection of target nucleic acids by forming a nucleic acid/photoreporter adduct layer on an ITO electrode. The target nucleic acids were hybridized with immobilized oligonucleotide capture probes on the ITO electrode. A subsequent binding of a photoreporter—a photoactive threading bis-intercalator consisting of two N,N′-bis(3-propyl-imidazole)-1,4,5,8-naphthalene diimides (PIND) linked by a Ru(bpy)22+ (bpy = 2,2′-bipyridine) complex (PIND–Ru–PIND)—allowed for photoelectrochemical detection of the target nucleic acids. The extremely low dissociation rate of the adduct and the highly reversible photoelectrochemical response under visible light illumination (490 nm) make it possible to conduct nucleic acid detection, with a sensitivity enhancement of four orders of magnitude over voltammetry. These results demonstrate for the first time the potential of photoelectrochemical biosensors for PCR-free ultrasensitive detection of nucleic acids.

[1]  E. Brown,et al.  Quantitative analysis of mRNA amplification by in vitro transcription. , 2001, Nucleic acids research.

[2]  N. Turro,et al.  Paradigms, supermolecules, electron transfer and chemistry at a distance. What's the problem? The science or the paradigm? , 1998, JBIC Journal of Biological Inorganic Chemistry.

[3]  W. Wilson,et al.  Interaction of aromatic imides with deoxyribonucleic acid. Spectrophotometric and viscometric studies. , 1982, Biochemistry.

[4]  M. Wasielewski,et al.  Dynamics of photoinduced charge transfer and hole transport in synthetic DNA hairpins. , 2001, Accounts of chemical research.

[5]  J. Rusling,et al.  Square wave voltammetric detection of chemical DNA damage with catalytic poly(4-vinylpyridine)-Ru(bpy)2(2+) films. , 2002, Analytical Chemistry.

[6]  S O Kelley,et al.  Femtosecond dynamics of DNA-mediated electron transfer. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[7]  H. Xie,et al.  Direct detection of DNA with an electrocatalytic threading intercalator. , 2005, Analytical chemistry.

[8]  Joel S. Silverman,et al.  Detection of Attomole Quantitites of DNA Targets on Gold Microelectrodes by Electrocatalytic Nucleobase Oxidation , 2003 .

[9]  E. Vollmer,et al.  Comparison of in-situ hybridization, direct and indirect in-situ PCR as well as tyramide signal amplification for the detection of HPV , 1999, Histochemistry and Cell Biology.

[10]  J. Barton,et al.  Effects of strand and directional asymmetry on base–base coupling and charge transfer in double-helical DNA , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[11]  F. Zhou,et al.  Selective Desorption of Alkanethiols in Mixed Self-Assembled Monolayers for Subsequent Oligonucleotide Attachment and DNA Hybridization , 2001 .

[12]  H. Xie,et al.  A nucleic acid biosensor for gene expression analysis in nanograms of mRNA. , 2004, Analytical chemistry.

[13]  J. Rusling,et al.  Voltammetric sensor for chemical toxicity using [Ru(bpy)2poly(4-vinylpyridine)10Cl)+ as catalyst in ultrathin films. DNA damage from methylating agents and an enzyme-generated epoxide. , 2003, Analytical chemistry.

[14]  Adam Heller,et al.  Enzyme-amplified amperometric detection of 3000 copies of DNA in a 10-microL droplet at 0.5 fM concentration. , 2003, Analytical chemistry.

[15]  P. Lizardi,et al.  Mutation detection and single-molecule counting using isothermal rolling-circle amplification , 1998, Nature Genetics.

[16]  P. Salvador,et al.  Analysis of the transient photocurrent-time behaviour of a sintered n-SrTiO3 electrode in water photoelectrolysis , 1984 .

[17]  Armistead,et al.  Modification of indium tin oxide electrodes with nucleic acids: detection of attomole quantities of immobilized DNA by electrocatalysis , 2000, Analytical chemistry.

[18]  H. Xie,et al.  Amperometric detection of nucleic acid at femtomolar levels with a nucleic acid/electrochemical activator bilayer on gold electrode. , 2004, Analytical chemistry.

[19]  A. Eckhardt,et al.  Probing biomolecule recognition with electron transfer: electrochemical sensors for DNA hybridization. , 1997, Bioconjugate chemistry.

[20]  B. Giese.  Long-distance charge transport in DNA: the hopping mechanism. , 2000, Accounts of chemical research.

[21]  G. Pesole,et al.  Guinea pig p53 mRNA: identification of new elements in coding and untranslated regions and their functional and evolutionary implications. , 1999, Genomics.

[22]  G. Schuster,et al.  Long-range charge transfer in DNA: transient structural distortions control the distance dependence. , 2000, Accounts of chemical research.

[23]  Itamar Willner,et al.  Dendritic amplification of DNA analysis by oligonucleotide-functionalized Au-nanoparticles , 2000 .

[24]  S. Kingsmore,et al.  Rolling circle amplification: a new approach to increase sensitivity for immunohistochemistry and flow cytometry. , 2001, The American journal of pathology.

[25]  Akimitsu Okamoto,et al.  Photostimulated hole transport through a DNA duplex immobilized on a gold electrode. , 2004, Journal of the American Chemical Society.

[26]  J. Barton,et al.  Direct chemical evidence for charge transfer between photoexcited 2-aminopurine and guanine in duplex DNA. , 2004, Journal of the American Chemical Society.

[27]  M. Urdea,et al.  A novel method for the rapid detection of specific nucleotide sequences in crude biological samples without blotting or radioactivity; application to the analysis of hepatitis B virus in human serum. , 1987, Gene.

[28]  Hans-Achim Wagenknecht.  Reductive electron transfer and transport of excess electrons in DNA. , 2003, Angewandte Chemie.

[29]  G. Pourtois,et al.  Pathways for photoinduced charge separation in DNA hairpins. , 2003, Journal of the American Chemical Society.

[30]  R. Lokey,et al.  A New Class of Polyintercalating Molecules , 1997 .

[31]  J. Rusling,et al.  Catalytic square-wave voltammetric detection of DNA with reversible metallopolymer-coated electrodes , 2001 .

[32]  S. Hotchandani,et al.  Photoelectric properties of ITO/CdS/chlorophyll a/Ag heterojunction solar cells , 1991 .

[33]  S. Eddy.  Non–coding RNA genes and the modern RNA world , 2001, Nature Reviews Genetics.

[34]  A. Steel,et al.  Electrochemical quantitation of DNA immobilized on gold. , 1998, Analytical chemistry.

[35]  T. C. Jenkins.  Optical absorbance and fluorescence techniques for measuring DNA-drug interactions. , 1997, Methods in molecular biology.

[36]  F. Caruso,et al.  Quartz crystal microbalance study of DNA immobilization and hybridization for nucleic Acid sensor development. , 1997, Analytical chemistry.

[37]  T. Majima,et al.  Charge separation in DNA via consecutive adenine hopping. , 2004, Journal of the American Chemical Society.

[38]  David A Russell,et al.  Electrochemical control of protein monolayers at indium tin oxide surfaces for the reagentless optical biosensing of nitric oxide. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[39]  R A Gibbs,et al.  Multiplex PCR: advantages, development, and applications. , 1994, PCR methods and applications.