Highly sensitive electrochemical impedance spectroscopic detection of DNA hybridization based on Au nano-CNT/PAN nano films. Talanta

Key Laboratory of Eco-chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, China.
Talanta (Impact Factor: 3.55). 02/2009; 77(3):1021-6. DOI: 10.1016/j.talanta.2008.07.058
Source: PubMed


A polyaniline nanofibers (PAN(nano))/carbon paste electrode (CPE) was prepared via dopping PAN(nano) in the carbon paste. The nanogold (Au(nano)) and carbon nanotubes (CNT) composite nanoparticles were bound on the surface of the PAN(nano)/CPE. The immobilization and hybridization of the DNA probe on the Au(nano)-CNT/PAN(nano) films were investigated with differential pulse voltammetry (DPV) and cyclic voltammetry (CV) using methylene blue (MB) as indicator, and electrochemical impedance spectroscopy (EIS) using [Fe(CN)(6)](3-/4-) as redox probe. The voltammetric peak currents of MB increased dramatically owing to the immobilization of the probe DNA on the Au(nano)-CNT/PAN(nano) films, and then decreased obviously owing to the hybridization of the DNA probe with the complementary single-stranded DNA (cDNA). The electron transfer resistance (R(et)) of the electrode surface increased after the immobilization of the probe DNA on the Au(nano)-CNT/PAN(nano) films and rose further after the hybridization of the probe DNA. The remarkable difference between the R(et) value at the DNA-immobilized electrode and that at the hybridized electrode could be used for the label-free EIS detection of the target DNA. The loading of the DNA probe on Au(nano)-CNT/PAN(nano) films was greatly enhanced and the sensitivity for the target DNA detection was markedly improved. The sequence-specific DNA of phosphinothricin acetyltransferase (PAT) gene and the polymerase chain reaction (PCR) amplification of nopaline synthase (NOS) gene from transgenically modified beans were determined with this label-free EIS DNA detection method. The dynamic range for detecting the PAT gene sequence was from 1.0 x 10(-12)mol/L to 1.0 x 10(-6)mol/L with a detection limit of 5.6 x 10(-13)mol/L.

13 Reads
  • Source
    • "The composite of conducting polymer-metal nanoparticle can be obtained from different metals and π-conjugated polymers as well as oligomer linkers which have received considerable attention due to the possibilities of creating suitable materials for electrocatalysis, chemical sensor and microelectronics [14,15]. Nanocomposite materials involving a hybrid of gold nanoparticle and carbon nanotube have been utilized for the construction of DNA biosensors [16]. Feng and his co-workers reported the use of a nanocomposite material consisting of gold nanoparticle and polyaniline nanotube membrane for DNA biosensor which was employed as a platform for immobilization of the DNA probe [2]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: A simple and highly sensitive electrochemical DNA aptasensor with high affinity for endocrine disrupting 17β-estradiol, was developed. Poly(3,4-ethylenedioxylthiophene) (PEDOT) doped with gold nanoparticles (AuNPs) was electrochemically synthesized and employed for the immobilization of biotinylated aptamer towards the detection of the target. The diffusion coefficient of the nanocomposite was 6.50 × 10(-7) cm(2) s(-1), which showed that the nanocomposite was highly conducting. Electrochemical impedance investigation also revealed the catalytic properties of the nanocomposite with an exchange current value of 2.16 × 10(-4) A, compared to 2.14 × 10(-5) A obtained for the bare electrode. Streptavidin was covalently attached to the platform using carbodiimide chemistry and the aptamer immobilized via streptavidin-biotin interaction. The electrochemical signal generated from the aptamer-target molecule interaction was monitored electrochemically using cyclic voltammetry and square wave voltammetry in the presence of [Fe(CN)(6)](-3/-4) as a redox probe. The signal observed shows a current decrease due to interference of the bound 17β-estradiol. The current drop was proportional to the concentration of 17β-estradiol. The PEDOT/AuNP platform exhibited high electroactivity, with increased peak current. The platform was found suitable for the immobilization of the DNAaptamer. The aptasensor was able to distinguish 17β-estradiol from structurally similar endocrine disrupting chemicals denoting its specificity to 17β-estradiol. The detectable concentration range of the 17β-estradiol was 0.1 nM-100 nM, with a detection limit of 0.02 nM.
    Full-text · Article · Nov 2010 · Sensors
  • Source
    • " using a 5′ thiol linker , has been used for detecting DNA hybridization ( Zhang et al . , 2009d ) . The response signal of interca - lated Adriamycin , corresponding to the DNA hybridization , is detected by differential pulse voltammetry . A highly sensitive technique based on impedance spectroscopy was reported for detecting DNA hybridization ( Zhou et al . , 2009 ) . The detection sensitivity for the PAT gene sequence is greatly increased by using nano Au - CNT / polyaniline nanofibers films for binding probe DNA . A rapid , reproducible and sensitive method is also developed for the biomolecular recognition of cis - diamminedichloroplatinum , an antican - cer drug using SWCNT modified disposabl"
    [Show abstract] [Hide abstract]
    ABSTRACT: Electrochemical (EC) sensing approaches have exploited the use of carbon nanotubes (CNTs) as electrode materials owing to their unique structures and properties to provide strong electrocatalytic activity with minimal surface fouling. Nanofabrication and device integration technologies have emerged along with significant advances in the synthesis, purification, conjugation and biofunctionalization of CNTs. Such combined efforts have contributed towards the rapid development of CNT-based sensors for a plethora of important analytes with improved detection sensitivity and selectivity. The use of CNTs opens an opportunity for the direct electron transfer between the enzyme and the active electrode area. Of particular interest are also excellent electrocatalytic activities of CNTs on the redox reaction of hydrogen peroxide and nicotinamide adenine dinucleotide, two major by-products of enzymatic reactions. This excellent electrocatalysis holds a promising future for the simple design and implementation of on-site biosensors for oxidases and dehydrogenases with enhanced selectivity. To date, the use of an anti-interference layer or an artificial electron mediator is critically needed to circumvent unwanted endogenous electroactive species. Such interfering species are effectively suppressed by using CNT based electrodes since the oxidation of NADH, thiols, hydrogen peroxide, etc. by CNTs can be performed at low potentials. Nevertheless, the major future challenges for the development of CNT-EC sensors include miniaturization, optimization and simplification of the procedure for fabricating CNT based electrodes with minimal non-specific binding, high sensitivity and rapid response followed by their extensive validation using "real world" samples. A high resistance to electrode fouling and selectivity are the two key pending issues for the application of CNT-based biosensors in clinical chemistry, food quality and control, waste water treatment and bioprocessing.
    Full-text · Article · Oct 2010 · Biotechnology advances
  • Source
    • "When the flat-band potential of the BDD was obtained from the Mott-Schottky plot, in which 1/C2 was plotted against the potential, they observed negative shifts in flat-band potentials upon hybridization with a complementary target, suggesting that the hybridization induce the decreased band bending of the p-type semiconductor. They further reported conducting polymer nanocomposites mixed with nanomaterials such as the carbon-nanotubes and TiO2 nanoparticles or nanotubes as a platform for DNA sensors for immobilizing the probe DNA [64-66]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Recent advances in label free DNA hybridization sensors employing electrochemical impedance spectroscopy (EIS) as a detection tool are reviewed. These sensors are based on the modulation of the blocking ability of an electrode modified with a probe DNA by an analyte, i.e., target DNA. The probe DNA is immobilized on a self-assembled monolayer, a conducting polymer film, or a layer of nanostructures on the electrode such that desired probe DNA would selectively hybridize with target DNA. The rate of charge transfer from the electrode thus modified to a redox indicator, e.g., [Fe(CN)6]3–/4–, which is measured by EIS in the form of charge transfer resistance (Rct), is modulated by whether or not, as well as how much, the intended target DNA is selectively hybridized. Efforts made to enhance the selectivity as well as the sensitivity of DNA sensors and to reduce the EIS measurement time are briefly described along with brief future perspectives in developing DNA sensors.
    Full-text · Article · Dec 2009 · Sensors
Show more