Highly sensitive electrochemiluminescence detection of single-nucleotide polymorphisms based on isothermal cycle-assisted triple-stem probe with dual-nanoparticle label.

We report here a new electrochemiluminescence (ECL) approach for detection of single nucleotide polymorphisms (SNPs) based on isothermal cycle-assisted triple-stem probe labeled with Au nanoparticles (NPs) and CdTe NPs. The system is composed of a CdS nanocrystals (NCs) film on glassy carbon electrode (GCE) as ECL emitter attached a double-stem DNA probe labeled with Au NPs. Then, the third stem labeled with CdTe NPs hybridizes with the double-stem DNA to form a triple-stem probe with the two labels near the CdS NCs film. A dual-quenched ECL of CdS NCs film is achieved due to energy transfer (ET) from CdS NCs to Au NPs and CdTe NPs, which makes the sensor exhibit relatively low background. Once the one base mutant DNA (mDNA) sequence as target of SNPs analysis displaces the third stem and hybridizes with the double-stem probe, forcing Au NPs away from the CdS NCs film, an ECL enhancement by the ECL-induced surface plasmon resonance of Au NPs is observed. Furthermore, after an isothermal cycle induced by primer, polymerase, and nicking endonuclease (NEase), a further enhancement of ECL is obtained. Taking advantages of the isothermal circular amplification system and the triple-stem probe architecture which enables turning its high selectivity toward specific target sequences, the reported biosensor shows excellent discrimination capabilities of SNPs with high selectivity and low detection limit (35 aM).

[1]  Jing‐Juan Xu,et al.  Ultrasensitive DNA detection based on Au nanoparticles and isothermal circular double-assisted electrochemiluminescence signal amplification. , 2011, Chemical communications.

[2]  Jing-Juan Xu,et al.  Enhanced electrochemiluminescence quenching of CdS:Mn nanocrystals by CdTe QDs-doped silica nanoparticles for ultrasensitive detection of thrombin. , 2011, Nanoscale.

[3]  Jing‐Juan Xu,et al.  CdS quantum dots/Ru(bpy)3(2+) electrochemiluminescence resonance energy transfer system for sensitive cytosensing. , 2011, Chemical communications.

[4]  Jing Wang,et al.  Gold nanoparticle enhanced electrochemiluminescence of CdS thin films for ultrasensitive thrombin detection. , 2011, Analytical chemistry.

[5]  Taihong Wang,et al.  Positive potential operation of a cathodic electrogenerated chemiluminescence immunosensor based on luminol and graphene for cancer biomarker detection. , 2011, Analytical chemistry.

[6]  Q. Guo,et al.  Entropy-driven molecular switch and signal amplification for homogeneous SNPs detection. , 2011, Chemical communications.

[7]  Taihong Wang,et al.  Highly sensitive electrogenerated chemiluminescence biosensor in profiling protein kinase activity and inhibition using gold nanoparticle as signal transduction probes. , 2010, Analytical chemistry.

[8]  Kemin Wang,et al.  Combination of DNA ligase reaction and gold nanoparticle-quenched fluorescent oligonucleotides: a simple and efficient approach for fluorescent assaying of single-nucleotide polymorphisms. , 2010, Analytical chemistry.

[9]  H. Ju,et al.  Quantum dots based electrochemiluminescent immunosensor by coupling enzymatic amplification with self-produced coreactant from oxygen reduction. , 2010, Analytical chemistry.

[10]  Jing‐Juan Xu,et al.  Electrochemiluminescence quenching by CdTe quantum dots through energy scavenging for ultrasensitive detection of antigen. , 2010, Chemical communications.

[11]  Hongyuan Chen,et al.  Opto-magnetic interaction between electrochemiluminescent CdS : Mn film and Fe3O4 nanoparticles and its application to immunosensing. , 2010, Chemical communications.

[12]  H. Ju,et al.  Low-potential electrochemiluminescent sensing based on surface unpassivation of CdTe quantum dots and competition of analyte cation to stabilizer. , 2010, Analytical chemistry.

[13]  Da Xing,et al.  Electrochemiluminescence biobarcode method based on cysteamine--gold nanoparticle conjugates. , 2010, Analytical chemistry.

[14]  Ronghua Yang,et al.  Rolling circle amplification combined with gold nanoparticle aggregates for highly sensitive identification of single-nucleotide polymorphisms. , 2010, Analytical chemistry.

[15]  A. Kotlyar,et al.  Synthesis and AFM characterization of poly(dG)-poly(dC)-gold nanoparticle conjugates. , 2010, Bioconjugate chemistry.

[16]  Chunhai Fan,et al.  Inhibition of the in vitro replication of DNA by an aptamer-protein complex in an autonomous DNA machine. , 2009, Chemistry.

[17]  H. Soh,et al.  An electrochemical sensor for single nucleotide polymorphism detection in serum based on a triple-stem DNA probe. , 2009, Journal of the American Chemical Society.

[18]  Kevin W Plaxco,et al.  Fluorescence detection of single-nucleotide polymorphisms with a single, self-complementary, triple-stem DNA probe. , 2009, Angewandte Chemie.

[19]  Jing-Juan Xu,et al.  Signal‐On Electrochemiluminescence Biosensors Based on CdS–Carbon Nanotube Nanocomposite for the Sensitive Detection of Choline and Acetylcholine , 2009 .

[20]  Jing-Juan Xu,et al.  Distance-dependent quenching and enhancing of electrochemiluminescence from a CdS:Mn nanocrystal film by Au nanoparticles for highly sensitive detection of DNA. , 2009, Chemical communications.

[21]  Kevin W Plaxco,et al.  Optimization of a reusable, DNA pseudoknot-based electrochemical sensor for sequence-specific DNA detection in blood serum. , 2009, Analytical chemistry.

[22]  Shusheng Zhang,et al.  Electrochemical DNA biosensor based on nanoporous gold electrode and multifunctional encoded DNA-Au bio bar codes. , 2008, Analytical chemistry.

[23]  H. Ju,et al.  Coreactant enhanced anodic electrochemiluminescence of CdTe quantum dots at low potential for sensitive biosensing amplified by enzymatic cycle. , 2008, Analytical chemistry.

[24]  Itamar Willner,et al.  Optical analysis of Hg2+ ions by oligonucleotide-gold-nanoparticle hybrids and DNA-based machines. , 2008, Angewandte Chemie.

[25]  Y. Wan,et al.  An enzyme-based E-DNA sensor for sequence-specific detection of femtomolar DNA targets. , 2008, Journal of the American Chemical Society.

[26]  Hongyuan Chen,et al.  Electrochemiluminescence immunosensor based on CdSe nanocomposites. , 2008, Analytical chemistry.

[27]  S. Dong,et al.  Quantum Dot Electrochemiluminescence in Aqueous Solution at Lower Potential and Its Sensing Application , 2008 .

[28]  D. Y. Zhang,et al.  Engineering Entropy-Driven Reactions and Networks Catalyzed by DNA , 2007, Science.

[29]  Wenwan Zhong,et al.  Typing of multiple single-nucleotide polymorphisms by a microsphere-based rolling circle amplification assay. , 2007, Analytical chemistry.

[30]  H. Ju,et al.  Anodic electrochemiluminescence of CdTe quantum dots and its energy transfer for detection of catechol derivatives. , 2007, Analytical chemistry.

[31]  A. Heeger,et al.  Label-free electrochemical detection of DNA in blood serum via target-induced resolution of an electrode-bound DNA pseudoknot. , 2007, Journal of the American Chemical Society.

[32]  H. Ju,et al.  Electrochemiluminescence sensors for scavengers of hydroxyl radical based on its annihilation in CdSe quantum dots film/peroxide system. , 2007, Analytical chemistry.

[33]  Hongyuan Chen,et al.  CdS nanocrystal-based electrochemiluminescence biosensor for the detection of low-density lipoprotein by increasing sensitivity with gold nanoparticle amplification. , 2007, Analytical chemistry.

[34]  Itamar Willner,et al.  Spotlighting of cocaine by an autonomous aptamer-based machine. , 2007, Journal of the American Chemical Society.

[35]  Itamar Willner,et al.  The aggregation of Au nanoparticles by an autonomous DNA machine detects viruses. , 2007, Small.

[36]  Hui Jiang,et al.  Enzyme-quantum dots architecture for highly sensitive electrochemiluminescence biosensing of oxidase substrates. , 2007, Chemical communications.

[37]  Itamar Willner,et al.  A virus spotlighted by an autonomous DNA machine. , 2006, Angewandte Chemie.

[38]  Huifeng Qian,et al.  A resonance energy transfer between chemiluminescent donors and luminescent quantum-dots as acceptors (CRET). , 2006, Angewandte Chemie.

[39]  A. Marx,et al.  Genotyping--from genomic DNA to genotype in a single tube. , 2005, Angewandte Chemie.

[40]  Tom Brown,et al.  Fluorescence based strategies for genetic analysis. , 2005, Chemical communications.

[41]  D. Clayton,et al.  Genome-wide association studies: theoretical and practical concerns , 2005, Nature Reviews Genetics.

[42]  M. Olivier.  A haplotype map of the human genome , 2003, Nature.

[43]  H. Ju,et al.  Electrogenerated chemiluminescence from a CdSe nanocrystal film and its sensing application in aqueous solution. , 2004, Analytical chemistry.

[44]  Catherine J. Murphy,et al.  Seed-Mediated Synthesis of Gold Nanorods: Role of the Size and Nature of the Seed , 2004 .

[45]  A. Bard,et al.  Electrochemistry and electrogenerated chemiluminescence of CdTe nanoparticles , 2004 .

[46]  A. Bard,et al.  Electrogenerated Chemiluminescence of CdSe Nanocrystals , 2002 .

[47]  N Risch,et al.  The Future of Genetic Studies of Complex Human Diseases , 1996, Science.