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Sample records for electrochemical dna biosensor

  1. Electrochemical application of DNA biosensors

    NASA Astrophysics Data System (ADS)

    Mascini, M.; Lucarelli, F.; Palchetti, I.; Marrazza, G.

    2001-09-01

    Disposable electrochemical DNA-based biosensors are reviewed; they have been used for the determination of low- molecular weight compounds with affinity for nucleic acids and for the detection of hybridization reaction. The first application is related to the molecular interaction between surface-linked DNA and pollutants or drugs, in order to develop a simple device for rapid screening of toxic compounds. The determination of such compounds was measured by their effect simple device for rapid screening of toxic compounds. The determination of such compounds was measured by their effect on the oxidation signal of the guanine peak of calf thymus DNA immobilized on the electrode surface and investigated by chronopotentiometric or voltammetric analysis. Applicability to river and wastewater sample is demonstrated. Moreover, disposable electrochemical sensors for the detection of a specific sequence of DNA were realized by immobilizing synthetic single-stranded oligonucleotides onto a graphite screen-printed electrode. The probes because hybridized with different concentrations of complementary sequences present in the sample. The hybrids formed on the electrode surface were evaluated by chronopotentiometric analysis using daunomycin as the indicator of the hybridization reaction. The hybridization was also performed using real samples. Application to apolipoprotein E is described, in this case samples have to be amplified by PCR and then analyzed by the DNA biosensor. The extension of such procedures to samples of environmental interest or to contamination of food is discussed.

  2. DNA-Metallodrugs Interactions Signaled by Electrochemical Biosensors: An Overview

    PubMed Central

    Ravera, Mauro; Bagni, Graziana; Mascini, Marco; Osella, Domenico

    2007-01-01

    The interaction of drugs with DNA is an important aspect in pharmacology. In recent years, many important technological advances have been made to develop new techniques to monitor biorecognition and biointeraction on solid devices. The interaction between DNA and drugs can cause chemical and conformational modifications and, thus, variation of the electrochemical properties of nucleobases. The propensity of a given compound to interact with DNA is measured as a function of the decrease of guanine oxidation signal on a DNA electrochemical biosensor. Covalent binding at N7 of guanine, electrostatic interactions, and intercalation are the events that this kind of biosensor can detect. In this context, the interaction between a panel of antitumoral Pt-, Ru-, and Ti-based metallodrugs with DNA immobilized on screen-printed electrodes has been studied. The DNA biosensors are used for semiquantitative evaluation of the analogous interaction occurring in the biological environment. PMID:18354727

  3. Electrochemical uranyl cation biosensor with DNA oligonucleotides as receptor layer.

    PubMed

    Jarczewska, Marta; Ziółkowski, Robert; Górski, Łukasz; Malinowska, Elżbieta

    2014-04-01

    The present study aims at the further development of the uranyl oligonucleotide-based voltammetric biosensor, which takes advantage of strong interaction between UO2(2+) and phosphate DNA backbone. Herein we report the optimization of working parameters of previously elaborated electrochemical DNA biosensor. It is shown that the sensor sensitivity is highly dependent on the oligonucleotide probe length and the incubation time of sensor in a sample solution. Consequently, the highest sensitivity was obtained for 10-nucleotide sequence and 60 min incubation time. The lower detection limit towards uranyl cation for developed biosensor was 30 nM. The influence of mixed monolayers and the possibility of developing a non-calibration device were also investigated. The selectivity of the proposed biosensor was significantly improved via elimination of adenine nucleobases from the DNA probe. Moreover, the regeneration procedure was elaborated and tested to prolong the use of the same biosensor for 4 subsequent determinations of UO2(2+). PMID:24334186

  4. Aminated hollow silica spheres for electrochemical DNA biosensor

    NASA Astrophysics Data System (ADS)

    Ariffin, Eda Yuhana; Heng, Lee Yook; Futra, Dedi; Ling, Tan Ling

    2015-09-01

    An electrochemical DNA biosensor for e.coli determination based on aminated hollow silica was successfully developed. Aminated hollow silica spheres were prepared through the reaction of Tween 20 template and silica precursor. The template was removed by the thermal decomposition at 620°C. Hollow silica spheres were modified with (3-Aminopropyl) triethoxysilane (APTS) to form aminated hollow silica spheres.Aminated DNA probe were covalently immobilized on to the amine functionalized hollow silica spheres through glutaradehyde linkers. The formation hollow silica was characterized using FTIR and FESEM. A range of 50-300nm particle size obtained from FESEM micrograph. Meanwhile for the electrochemical study, a quasi-reversible system has been obtain via cyclic voltammetry (CV).

  5. Ultrasensitive electrochemical cocaine biosensor based on reversible DNA nanostructure.

    PubMed

    Sheng, Qinglin; Liu, Ruixiao; Zhang, Sai; Zheng, Jianbin

    2014-01-15

    We proposed an ultrasensitive electrochemical cocaine biosensor based on the three-dimensional (3D) DNA structure conversion of nanostructure from Triangular Pyramid Frustum (TPFDNA) to Equilateral Triangle (ETDNA). The presence of cocaine triggered the aptamer-composed DNA nanostructure change from "Close" to "Open", leading to obvious faradaic impedance changes. The unique properties with excellent stability and specific rigid structure of the 3D DNA nanostructure made the biosensing functions stable, sensitive, and regenerable. The Faradaic impedance responses were linearly related to cocaine concentration between 1.0 nM and 2.0 μM with a correlation coefficient of 0.993. The limit of detection was calculated to be 0.21 nM following IUPAC recommendations (3Sb/b). It is expected that the distinctive features of DNA nanostructure would make it potentially advantageous for a broad range of biosensing, bionanoelectronics, and therapeutic applications. PMID:23962705

  6. Electrochemical DNA biosensor based on the BDD nanograss array electrode

    PubMed Central

    2013-01-01

    Background The development of DNA biosensor has attracted considerable attention due to their potential applications, including gene analysis, clinical diagnostics, forensic study and more medical applications. Using electroactive daunomycin as an indicator, the hybridization detection was measured by differential pulse voltammetry in this study. Results Electrochemical DNA biosensor was developed based on the BDD film electrode (fBDD) and BDD nanograss array electrode (nBDD). In comparison with fBDD and AuNPs/CA/fBDD electrode, the lower semicircle diameter of electrochemical impedance spectroscopy obtained on nBDD and AuNPs/CA/nBDD electrode indicated that the presence of nanograss array improved the reactive site, reduced the interfacial resistance, and made the electron transfer easier. Using electroactive daunomycin as an indicator, the hybridization detection was measured by differential pulse voltammetry. Conclusions The experimental results demonstrated that the prepared AuNPs/CA/nBDD electrode was suitable for DNA hybridization with favorable performance of faster response, higher sensitivity, lower detection limit and satisfactory selectivity, reproducibility and stability. PMID:23575250

  7. Genomagnetic Electrochemical Biosensors

    NASA Astrophysics Data System (ADS)

    Wang, Joseph; Erdem, Arzum

    The use of nucleic acid technologies has significantly improved preparation and diagnostic procedures in life sciences. Nucleic acid layers combined with electrochemical or optical transducers produce a new kind of affinity biosensors as DNA Biosensor for small molecular weight molecules. Electrochemical DNA biosensors are attractive devices for converting the hybridization event into an analytical signal for obtaining sequence-specific information in connection with clinical, environmental or forensic investigations. DNA hybridization biosensors, based on electrochemical transduction of hybridization, couple the high specificity of hybridization reactions with the excellent sensitivity and portability of electrochemical transducers. The main goal in all researches is to design DNA biosensors for preparing a basis for the future DNA microarray system. DNA chip has now become a powerful tool in biological research, however the real clinic assay is still under development. Recently, there has been a great interest to the magnetic beads and/or nanoparticles labelled with metals such as gold, cadmium, silver, etc. for designing of novel electrochemical DNA biosensor approaches resulting in efficient separation. The attractive features of this technology include simple approach, rapid results, multi-analyte detection, low-cost per measurument, stable, and non-hazardous reagents, and reduced waste handling. Some of these new approaches and applications of the electrochemical DNA biosensors based on magnetic beads and its combining with nanoparticles labelled with metals are described and discussed.

  8. [Cu(phen)2](2+) acts as electrochemical indicator and anchor to immobilize probe DNA in electrochemical DNA biosensor.

    PubMed

    Yang, Linlin; Li, Xiaoyu; Li, Xi; Yan, Songling; Ren, Yinna; Wang, Mengmeng; Liu, Peng; Dong, Yulin; Zhang, Chaocan

    2016-01-01

    We demonstrate a novel protocol for sensitive in situ label-free electrochemical detection of DNA hybridization based on copper complex ([Cu(phen)2](2+), where phen = 1,10-phenanthroline) and graphene (GR) modified glassy carbon electrode. Here, [Cu(phen)2](2+) acted advantageously as both the electrochemical indicator and the anchor for probe DNA immobilization via intercalative interactions between the partial double helix structure of probe DNA and the vertical aromatic groups of phen. GR provided large density of docking site for probe DNA immobilization and increased the electrical conductivity ability of the electrode. The modification procedure was monitored by electrochemical impedance spectroscopy (EIS). Square-wave voltammetry (SWV) was used to explore the hybridization events. Under the optimal conditions, the designed electrochemical DNA biosensor could effectively distinguish different mismatch degrees of complementary DNA from one-base mismatch to noncomplementary, indicating that the biosensor had high selectivity. It also exhibited a reasonable linear relationship. The oxidation peak currents of [Cu(phen)2](2+) were linear with the logarithm of the concentrations of complementary target DNA ranging from 1 × 10(-12) to 1 × 10(-6) M with a detection limit of 1.99 × 10(-13) M (signal/noise = 3). Moreover, the stability of the electrochemical DNA biosensor was also studied. PMID:26403602

  9. Electrochemical DNA biosensor based on avidin-biotin conjugation for influenza virus (type A) detection

    NASA Astrophysics Data System (ADS)

    Chung, Da-Jung; Kim, Ki-Chul; Choi, Seong-Ho

    2011-09-01

    An electrochemical DNA biosensor (E-DNA biosensor) was fabricated by avidin-biotin conjugation of a biotinylated probe DNA, 5'-biotin-ATG AGT CTT CTA ACC GAG GTC GAA-3', and an avidin-modified glassy carbon electrode (GCE) to detect the influenza virus (type A). An avidin-modified GCE was prepared by the reaction of avidin and a carboxylic acid-modified GCE, which was synthesized by the electrochemical reduction of 4-carboxyphenyl diazonium salt. The current value of the E-DNA biosensor was evaluated after hybridization of the probe DNA and target DNA using cyclic voltammetry (CV). The current value decreased after the hybridization of the probe DNA and target DNA. The DNA that was used follows: complementary target DNA, 5'-TTC GAC CTC GGT TAG AAG ACT CAT-3' and two-base mismatched DNA, 5'-TTC GAC AGC GGT TAT AAG ACT CAT-3'.

  10. DNA electrochemical biosensor for metallic drugs at physiological conditions

    PubMed Central

    Santiago-Lopez, Angel J.; Vera, José L.; Meléndez, Enrique

    2014-01-01

    Entrapment of dsSS-DNA into the polypyrrole-polyvinyl sulphonate (dsSS-DNA-PPy-PVS) film over indium-tin-oxide (ITO) coated glass has been designed to detect titanium and platinum drugs, titanocene dichloride and cisplatin. The disposable dsSS-DNA-PPy-PVS/ITO biosensor was characterized by cyclic voltammetry, attenuated total reflectance Infrared spectroscopy and atomic force microscopy. Amperometric studies by cyclic voltammetry using, dsSS-DNA-PPy PVS/ITO biosensor, demonstrated the ability of this biosensor to detect these metallic drugs in millimolar concentration by monitoring the decrease of the guanine oxidation signal as a result of the DNA damage. The concentration range detected for titanocene dichloride is 0.25 to 1.5 mM and for cisplatin is 0.06 to 1.0 mM. PMID:25705144

  11. DNA-gold nanoparticles network based electrochemical biosensors for DNA MTase activity.

    PubMed

    Hong, Lu; Wan, Jing; Zhang, Xiaojun; Wang, Guangfeng

    2016-05-15

    In this work, a highly sensitive electrochemical DNA methyltransferase (MTase) activity assay was fabricated with DNA-gold nanoparticles (Au NPs) network as signal amplification unit and an easy assembly method by the linkage of benzenedithiol bridge. By two complementary AuNPs modified single-stranded DNA, DNA-gold nanoparticles network was self-assembled. With the linkage of benzenedithiol bridge, the DNA network structure was immobilized on the surface of gold electrode through the covalent Au-S bond. In the presence of Dam MTase, the special sites of DNA-AuNPs network were methylated and could not be digested by restriction endonuclease Mbo I. Thus the loaded electrochemical indicator Methylene blue (MB) was MB molecules still remained on the DNA-Au NPs network. The electrochemical response depended on the methylated degree, which could be used to detect MTase activity. By the differential pulse voltammetry (DPV), it was demonstrated that a linear relationship between the DPV response and logarithm of Dam concentration ranged from 0.075 to 30U/mL, achieving a low detection limit of 0.02U/mL. The use of benzenedithiol avoided the direct incubation of the solid electrode with the capture DNA probe under complex and harsh conditions. Therefore the immobilization of DNA-AuNPs network was easy to be carried out, which is favorable for the specially high stability and reproducibility of the electrochemical biosensor. PMID:26992515

  12. Gelatin methacrylate (GelMA) mediated electrochemical DNA biosensor for DNA hybridization.

    PubMed

    Topkaya, Seda Nur

    2015-02-15

    In this study, an electrochemical biosensor system for the detection of DNA hybridization by using gelatin methacrylate (GelMA) modified electrodes was developed. Electrochemical behavior of GelMA modified Pencil Graphite Electrode (PGE) that serve as a functional platform was investigated by using Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS) and compared with those of the bare PGE. Hybridization was achieved in solution phase and guanine oxidation signal changes were evaluated. The decrease in the guanine oxidation peak currents at around +1.0 V was used as an indicator for the DNA hybridization. Also, more interestingly GelMA intrinsic oxidation peaks at around +0.7 V changed substantially by immobilization of different oligonucleotides such as probe, hybrid and control sequences to the electrode surface. It is the first study of using GelMA as a part of an electrochemical biosensor system. The results are very promising in terms of using GelMA as a new DNA hybridization indicator. Additionally, GelMA modified electrodes could be useful for detecting ultra low quantity of oligonucleotides by providing mechanical support to the bio-recognition layer. The detection limit of this method is at present 10(-12)mol. Signal suppressions were increased from 50% to 93% for hybrid with using GelMA when it was compared to bare electrode which facilitates the hybridization detection. PMID:25286352

  13. Characterizing metabolic inhibition using electrochemical enzyme/DNA biosensors.

    PubMed

    Hull, Dominic O; Bajrami, Besnik; Jansson, Ingela; Schenkman, John B; Rusling, James F

    2009-01-15

    Studies of metabolic enzyme inhibition are necessary in drug development and toxicity investigations as potential tools to limit or prevent appearance of deleterious metabolites formed, for example, by cytochrome (cyt) P450 enzymes. In this paper, we evaluate the use of enzyme/DNA toxicity biosensors as tools to investigate enzyme inhibition. We have examined DNA damage due to cyt P450cam metabolism of styrene using DNA/enzyme films on pyrolytic graphite (PG) electrodes monitored via Ru(bpy)(3)(2+)-mediated DNA oxidation. Styrene metabolism initiated by hydrogen peroxide was evaluated with and without the inhibitors, imidazole, imidazole-4-acetic acid, and sulconazole (in micromolar range) to monitor DNA damage inhibition. The initial rates of DNA damage decreased with increased inhibitor concentrations. Linear and nonlinear fits of Michaelis-Menten inhibition models were used to determine apparent inhibition constants (K(I)*) for the inhibitors. Elucidation of the best fitting inhibition model was achieved by comparing correlation coefficients and the sum of the square of the errors (SSE) from each inhibition model. Results confirmed the utility of the enzyme/DNA biosensor for metabolic inhibition studies. A simple competitive inhibition model best approximated the data for imidazole, imidazole-4-acetic acid and sulconazole with K(I)* of 268.2, 142.3, and 204.2 microM, respectively. PMID:19099359

  14. Characterizing Metabolic Inhibition Using Electrochemical Enzyme-DNA Biosensors

    PubMed Central

    Hull, Dominic O.; Bajrami, Besnik; Jansson, Ingela; Schenkman, John B.; Rusling, James F.

    2009-01-01

    Studies of metabolic enzyme inhibition are necessary in drug development and toxicity investigations as potential tools to limit or prevent appearance of deleterious metabolites formed, for example by cytochrome (cyt) P450 enzymes. In this paper, we evaluate the use of enzyme/DNA toxicity biosensors as tools to investigate enzyme inhibition. We have examined DNA damage due to cyt P450cam metabolism of styrene using DNA/enzyme films on pyrolytic graphite (PG) electro*des monitored via Ru(bpy)32+–mediated DNA oxidation. Styrene metabolism initiated by hydrogen peroxide was evaluated with and without the inhibitors, imidazole, imidazole-4-acetic acid and sulconazole (in micromolar range) to monitor DNA damage inhibition. The initial rates of DNA damage decreased with increased inhibitor concentrations. Linear and nonlinear fits of Michaelis-Menten inhibition models were used to determine apparent inhibition constants (KI*) for the inhibitors. Elucidation of the best fitting inhibition model was achieved by comparing correlation coefficients and the sum of the square of the errors (SSE) from each inhibition model. Results confirmed the utility of the enzyme/DNA biosensor for metabolic inhibition studies. A simple competitive inhibition model best approximated the data for imidazole, imidazole-4-acetic acid and sulconazole with KI* of 268.2, 142.3 and 204.2 µM, respectively. PMID:19099359

  15. Detection EGFR exon 19 status of lung cancer patients by DNA electrochemical biosensor.

    PubMed

    Xu, Xiong-Wei; Weng, Xiu-Hua; Wang, Chang-Lian; Lin, Wei-Wei; Liu, Ai-Lin; Chen, Wei; Lin, Xin-Hua

    2016-06-15

    Epidermal growth factor receptor (EGFR) exon 19 mutation status is a very important prediction index for tyrosine kinase inhibitors (TKIs) therapy. In this paper, we constructed a superior selective sandwich-type electrochemical biosensor to detect in-frame deletions in exon 19 of EGFR in real samples of patients with non-small cell lung carcinoma. Based on the characteristics of different hybridization efficiency in different hybridization phase conditions, different region around EGFR exon 19 deletion hotspots was selected to design DNA probes to improve biosensor performance. The results confirm that alteration of deletion location in target deliberately according to different hybridization phase is able to improve selectivity of sandwich-type DNA biosensor. Satisfactory discrimination ability can be achieved when the deletions are located in the capture probe interaction region. In order to improve efficiency of ssDNA generation from dsDNA, we introduce Lambda exonuclease (λ-exo) to sandwich-type biosensor system. EGFR exon 19 statuses of clinical real samples from lung cancer patients can be discriminated successfully by the proposed method. Our research would make the electrochemical biosensor be an excellent candidate for EGFR detection for lung cancer patients. PMID:26874108

  16. Ultrasensitive cDNA Detection of Dengue Virus RNA Using Electrochemical Nanoporous Membrane-Based Biosensor

    PubMed Central

    Rai, Varun; Hapuarachchi, Hapuarachchige C.; Ng, Lee Ching; Soh, Siew Hwa; Leo, Yee Sin; Toh, Chee-Seng

    2012-01-01

    A nanoporous alumina membrane-based ultrasensitive DNA biosensor is constructed using 5′-aminated DNA probes immobilized onto the alumina channel walls. Alumina nanoporous membrane-like structure is carved over platinum wire electrode of 76 µm diameter dimension by electrochemical anodization. The hybridization of complementary target DNA with probe DNA molecules attached inside the pores influences the pore size and ionic conductivity. The biosensor demonstrates linear range over 6 order of magnitude with ultrasensitive detection limit of 9.55×10−12 M for the quantification of ss-31 mer DNA sequence. Its applicability is challenged against real time cDNA PCR sample of dengue virus serotype1 derived from asymmetric PCR. Excellent specificity down to one nucleotide mismatch in target DNA sample of DENV3 is also demonstrated. PMID:22927927

  17. Nanobiomaterials for Electrochemical Biosensors

    NASA Astrophysics Data System (ADS)

    Pumera, M.

    2007-08-01

    I will discuss main techniques and methods which use nanoscale materials for construction of electrochemical biosensors with emphasis on methods developed by myself and my coworkers. Described approaches include carbon nanotube based electrodes relying on double wall and multiwall carbon nanotubes, novel binding materials and mass production technology; and nanoscale materials as biomolecule tracers, including gold nanoparticles for DNA detection. Specific issues related to electrochemistry of nanoscale materials will be discussed. Various applications for genomic and proteomic analysis will be described.

  18. Short thio-multi-walled carbon nanotubes and Au nanoparticles enhanced electrochemical DNA biosensor for DNA hybridization detection

    NASA Astrophysics Data System (ADS)

    Guo, Feng; Zhang, Jimei; Dai, Zhao; Zheng, Guo

    2010-07-01

    A novel and sensitive electrochemical DNA biosensor based on multi-walled carbon nanotubes functionalized with a thio group (MWNTs-SH) and gold nanoparticles (GNPs) for covalent DNA immobilization and enhanced hybridization detection is described. The key step for developing this novel DNA biosensor is to cut the pristine MWNT into short and generate lots of active sites simultaneously. With this approach, the target DNA could be quantified in a linear range from 8.5×10-10 to 1.5×10-5 mol/L, with a detection limit of 1.67×10-11 mol/L by 3σ.

  19. Novel electrochemical DNA hybridization biosensors for selective determination of silver ions.

    PubMed

    Ebrahimi, Maryam; Raoof, Jahan Bakhsh; Ojani, Reza

    2015-11-01

    In this work, novel electrochemical biosensors for Ag(+) determination based on Ag(+)-induced DNA hybridization, using Ethyl green (EG) as an electroactive label on the surface of bare carbon paste electrode (CPE) and gold nanoparticles-modified carbon paste electrode (GN-CPE) are reported. Two single-strand poly-C (100% cytosine bases) DNAs are used as oligonucleotide probe and target. In the presence of Ag(+), the target DNA with full cytosine-cytosine (C-C) mismatches could hybridize with the probe DNA by forming C-Ag(+)-C complex. The induced hybridization of the two oligonucleotides leads to the decrease in the reduction peak currents of EG, which could be used for electrochemical determination of Ag(+). This difference in the values of the reduction peak current of EG before and after DNA hybridization (∆I) was linear with the concentration of Ag(+) in the ranges from 3.0×10(-10) to 1.0×10(-9) mol L(-1) and 9.0×10(-11) to 1.0×10(-9) mol L(-1), for the biosensor and nanoparticles modified-biosensor, respectively. Calculated detection limits were 1.04×10(-10) and 2.64×10(-11) mol L(-1) for biosensor and nanoparticles modified-biosensor, respectively. The biosensors demonstrated good selectivity towards Ag(+) ions in the presence of some metal ions such as Pb(2+), Cu(2+), Ca(2+), Zn(2+), Fe(2+) and Hg(2+). The proposed biosensors were applied successfully to the voltammetric determination of Ag(+) in real samples. PMID:26452869

  20. Highly sensitive electrochemical biosensor based on nonlinear hybridization chain reaction for DNA detection.

    PubMed

    Jia, Liping; Shi, Shanshan; Ma, Rongna; Jia, Wenli; Wang, Huaisheng

    2016-06-15

    In the present work we demonstrated an ultrasensitive detection platform for specific DNA based on nonlinear hybridization chain reaction (HCR) by triggering chain-branching growth of DNA dendrimers. HCR was initiated by target DNA (tDNA) and finally formed dendritic structure by self-assembly. The electrochemical signal was drastically enhanced by capturing multiple catalytic peroxidase with high-ordered growth. Electrochemical signals were obtained by measuring the reduction current of oxidized 3, 3', 5, 5'-tetramethylbenzidine sulfate (TMB), which was generated by HRP in the presence of H2O2. This method exhibited ultrahigh sensitivity to tDNA with detection limit of 0.4fM. Furthermore, the biosensor was also capable of discriminating single-nucleotide difference among concomitant DNA sequences. PMID:26872213

  1. Dopamine-loaded liposome and its application in electrochemical DNA biosensor.

    PubMed

    Mahmoudi-Badiki, Tohid; Alipour, Esmaeel; Hamishehkar, Hamed; Golabi, Seyed Mahdi

    2016-08-01

    In this study, disruption and lyophilization-rehydration of dopamine-loaded liposome and its application in electrochemical DNA biosensor was investigated. The liposomes containing soyphosphatidylcholine and cholesterol were prepared through thin-layer hydration. First, an investigation was carried out to find an appropriate lysing agent for disruption of prepared liposomes. Differential pulse voltammetry, as a high sensitive electrochemical technique, was used along with a multi-walled carbon nanotubes modified glassy carbon electrode for sensitive electrochemical detection of released dopamine from disrupted liposomes. Various lysing agents were investigated and finally, the disruption of liposomes using methanol was selected without any surfactant, because of its least fouling effect. Then, lyophilization of dopamine-loaded liposomes was carried out using sucrose as cryoprotectant. The electrochemical studies of lyophilized liposomes showed that the remained dopamine in sucrose-protected liposomes was higher than sucrose-free liposomes. Furthermore, sucrose has no interference in electrochemical studies. Then, with the addition of biotin-X-DHPE to liposome formulation, the lyophilized sucrose protected dopamine-loaded biotin-tagged liposomes were prepared and the feasibility of application of them in electrochemical DNA biosensor was investigated as signal enhancer and verified for detection of oligonucleotides. PMID:27194602

  2. Identification of Chinese Herbs Using a Sequencing-Free Nanostructured Electrochemical DNA Biosensor

    PubMed Central

    Lei, Yan; Yang, Fan; Tang, Lina; Chen, Keli; Zhang, Guo-Jun

    2015-01-01

    Due to the nearly identical phenotypes and chemical constituents, it is often very challenging to accurately differentiate diverse species of a Chinese herbal genus. Although technologies including DNA barcoding have been introduced to help address this problem, they are generally time-consuming and require expensive sequencing. Herein, we present a simple sequencing-free electrochemical biosensor, which enables easy differentiation between two closely related Fritillaria species. To improve its differentiation capability using trace amounts of DNA sample available from herbal extracts, a stepwise electrochemical deposition of reduced graphene oxide (RGO) and gold nanoparticles (AuNPs) was adopted to engineer a synergistic nanostructured sensing interface. By using such a nanofeatured electrochemical DNA (E-DNA) biosensor, two Chinese herbal species of Fritillaria (F. thunbergii and F. cirrhosa) were successfully discriminated at the DNA level, because a fragment of 16-mer sequence at the spacer region of the 5S-rRNA only exists in F. thunbergii. This E-DNA sensor was capable of identifying the target sequence in the range from 100 fM to 10 nM, and a detection limit as low as 11.7 fM (S/N = 3) was obtained. Importantly, this sensor was applied to detect the unique fragment of the PCR products amplified from F. thunbergii and F. cirrhosa, respectively. We anticipate that such a direct, sequencing-free sensing mode will ultimately pave the way towards a new generation of herb-identification strategies. PMID:26633399

  3. Coupling electrochemical response of a DNA biosensor with PCR for Neisseria gonorrhoeae detection.

    PubMed

    Verma, Rachna; Sood, Seema; Singh, Renu; Sumana, Gajjala; Bala, Manju; Sharma, Vinod K; Samantaray, Jyotish C; Pandey, Ravindra M; Malhotra, Bansi D

    2014-01-01

    Early diagnosis of gonococcal infections is important with regard to a patient's health and stage of infection. In this context, we report the development of an opa-gene-based electrochemical DNA biosensor for detection of Neisseria gonorrhoeae by monitoring redox peak of methylene blue indicator. The fabricated biosensor has been shown to be highly sensitive and specific when evaluated with complementary, non-complementary, and 1-base mismatch DNA sequences and polymerase chain reaction (PCR) amplified products (amplicons) of standard strain of N. gonorrhoeae (ATCC49226). The biosensor has been further evaluated using amplicons of known positive and negative clinical samples, and cut-off for positives has been determined using receiver operating characteristic curve. The sensitivity (SN), specificity (SP), positive predictive value, and negative predictive value of the biosensor have been found to be 96.2%, 88.2%, 92.6%, and 93.8%, respectively. We conclude that the combination of PCR amplification with electrochemical detection shows distinct advantage of high SN and increased SP for gonococcal detection. PMID:24207077

  4. An Electrochemical DNA Biosensor for the Detection of Salmonella Using Polymeric Films and Electrochemical Labels

    NASA Astrophysics Data System (ADS)

    Diaz Serrano, Madeline

    Waterborne and foodborne diseases are one of the principal public health problems worldwide. Microorganisms are the major agents of foodborne illness: pathogens such as Salmonella, Campylobacter jejuni and Escherichia coli, and parasites such as cryptosporidium. The most popular methods to detect Salmonella are based on culture and colony counting methods, ELISA, Gel electrophoresis and the polymerase chain reaction. Conventional detection methods are laborious and time-consuming, allowing for portions of the food to be distributed, marketed, sold and eaten before the analysis is done and the problem even detected. By these reasons, the rapid, easy and portable detection of foodborne organisms will facilitate the disease treatment. Our particular interest is to develop a nucleic acid biosensor (NAB) for the detection of pathogenic microorganisms in food and water samples. In this research, we report on the development of a NAB prototype using a polymer modified electrode surface together with sequences of different lengths for the OmpC gene from Salmonella as probes and Ferrocene-labeled target (Fc-ssDNA), Ferrocene-labeled tri(ethylene glycol) (Fc-PEG) and Ruthenium-Ferrocene (Ru-Fe) bimetallic complex as an electrochemical labels. We have optimized several PS films and anchored nucleic acid sequences with different lengths at gold and carbon surfaces. Non contact mode AFM and XPS were used to monitor each step of the NAB preparation, from polymer modification to oligos hybridization (conventional design). The hybridization reaction was followed electrochemically using a Fc-ssDNA and Fc-PEG in solution taking advantage of the morphological changes generated upon hybridization. We observed a small current at the potential for the Fe oxidation without signal amplification at +296 mV vs. Ag/AgCl for the Fc-ssDNA strategy and a small current at +524 mV for the Fc-PEG strategy. The immobilization, hybridization and signal amplification of Biotin- OmpC Salmonella genes

  5. Nanostructured SERS-electrochemical biosensors for testing of anticancer drug interactions with DNA.

    PubMed

    Ilkhani, Hoda; Hughes, Taylor; Li, Jing; Zhong, Chuan Jian; Hepel, Maria

    2016-06-15

    Widely used anti-cancer treatments involving chemotherapeutic drugs result in cancer cell damage due to their strong interaction with DNA. In this work, we have developed laboratory biosensors for screening chemotherapeutic drugs and to aid in the assessment of DNA modification/damage caused by these drugs. The sensors utilize surface-enhanced Raman scattering (SERS) spectroscopy and electrochemical methods to monitor sensory film modification and observe the drug-DNA reactivity. The self-assembled monolayer protected gold-disk electrode (AuDE) was coated with a reduced graphene oxide (rGO), decorated with plasmonic gold-coated Fe2Ni@Au magnetic nanoparticles functionalized with double-stranded DNA (dsDNA), a sequence of the breast cancer gene BRCA1. The nanobiosensors AuDE/SAM/rGO/Fe2Ni@Au/dsDNA were then subjected to the action of a model chemotherapeutic drug, doxorubicin (DOX), to assess the DNA modification and its dose dependence. The designed novel nanobiosensors offer SERS/electrochemical transduction, enabling chemically specific and highly sensitive analytical signals generation. The SERS measurements have corroborated the DOX intercalation into the DNA duplex whereas the electrochemical scans have indicated that the DNA modification by DOX proceeds in a concentration dependent manner, with limit of detection LOD=8µg/mL (S/N=3), with semilog linearity over 3 orders of magnitude. These new biosensors are sensitive to agents that interact with DNA and facilitate the analysis of functional groups for determination of the binding mode. The proposed nanobiosensors can be applied in the first stage of the drug development for testing the interactions of new drugs with DNA before the drug efficacy can be assessed in more expensive testing in vitro and in vivo. PMID:26851584

  6. Electrochemical determination of biophenol oleuropein using a simple label-free DNA biosensor.

    PubMed

    Mohamadi, Maryam; Mostafavi, Ali; Torkzadeh-Mahani, Masoud

    2015-02-01

    Oleuropein (Ole), naturally occurring phenolic compound found in olive products, is well known for its benefits for human health. In the present work, a simple, sensitive and rapid determination of Ole was achieved using a label-free electrochemical DNA biosensor. The application was related to the molecular interaction between Ole and double-stranded DNA (dsDNA). So, the voltammetric behavior of Ole at the surface of a DNA-immobilized chitosan-modified carbon paste electrode was studied using differential pulse voltammetry (DPV) where the oxidation peak current of Ole was measured as an analytical signal. A considerable increase was observed in the oxidation signal of Ole at the DNA-coated electrode compared with the DNA-free electrode, indicating the pre-concentration of Ole due to the interaction with the surface-confined DNA layer. In order to use the proposed sensor for real samples, different parameters affecting Ole signal such as, immobilization time and potential, accumulation time and pH, and stripping pH were optimized. Under optimized experimental conditions, a linear concentration range of 0.30-12μmolL(-1) with a detection limit of 0.090μmolL(-1) was obtained for Ole determination. The proposed biosensor was successfully applied to the determination of Ole in olive leaf extract and human serum samples. PMID:25087151

  7. An ultrasensitive electrochemical DNA biosensor based on a copper oxide nanowires/single-walled carbon nanotubes nanocomposite

    NASA Astrophysics Data System (ADS)

    Chen, Mei; Hou, Changjun; Huo, Danqun; Yang, Mei; Fa, Huanbao

    2016-02-01

    Here, we developed a novel and sensitive electrochemical biosensor to detect specific-sequence target DNA. The biosensor was based on a hybrid nanocomposite consisting of copper oxide nanowires (CuO NWs) and carboxyl-functionalized single-walled carbon nanotubes (SWCNTs-COOH). The resulting CuO NWs/SWCNTs layers exhibited a good differential pulse voltammetry (DPV) current response for the target DNA sequences, which we attributed to the properties of CuO NWs and SWCNTs. CuO NWs and SWCNTs hybrid composites with highly conductive and biocompatible nanostructure were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and cyclic voltammetry (CV). Immobilization of the probe DNA on the electrode surface was largely improved due to the unique synergetic effect of CuO NWs and SWCNTs. DPV was applied to monitor the DNA hybridization event, using adriamycin as an electrochemical indicator. Under optimal conditions, the peak currents of adriamycin were linear with the logarithm of target DNA concentrations (ranging from 1.0 × 10-14 to 1.0 × 10-8 M), with a detection limit of 3.5 × 10-15 M (signal/noise ratio of 3). The biosensor also showed high selectivity to single-base mismatched target DNA. Compared with other electrochemical DNA biosensors, we showed that the proposed biosensor is simple to implement, with good stability and high sensitivity.

  8. Electrochemical biosensor modified with dsDNA monolayer for restriction enzyme activity determination.

    PubMed

    Zajda, Joanna; Górski, Łukasz; Malinowska, Elżbieta

    2016-06-01

    A simple and cost effective method for the determination of restriction endonuclease activity is presented. dsDNA immobilized at a gold electrode surface is used as the enzymatic substrate, and an external cationic redox probe is employed in voltammetric measurements for analytical signal generation. The assessment of enzyme activity is based on a decrease of a current signal derived from reduction of methylene blue which is present in the sample solution. For this reason, the covalent attachment of the label molecule is not required which significantly reduces costs of the analysis and simplifies the entire determination procedure. The influence of buffer components on utilized dsDNA/MCH monolayer stability and integrity is also verified. Electrochemical impedance spectroscopy measurements reveal that due to pinhole formation during enzyme activity measurement the presence of any surfactants should be avoided. Additionally, it is shown that the sensitivity of the electrochemical biosensor can be tuned by changing the restriction site location along the DNA length. Under optimal conditions the proposed biosensor exhibits a linear response toward PvuII activity within a range from 0.25 to 1.50 U/μL. PMID:26859430

  9. Recent Updates of DNA Incorporated in Carbon Nanotubes and Nanoparticles for Electrochemical Sensors and Biosensors

    PubMed Central

    Yogeswaran, Umasankar; Thiagarajan, Soundappan; Chen, Shen-Ming

    2008-01-01

    Innovations in the field of electrochemical sensors and biosensors are of much importance nowadays. These devices are designed with probes and micro electrodes. The miniaturized designs of these sensors allow analyses of materials without damaging the samples. Some of these sensors are also useful for real time analysis within the host system, so these sensors are considered to be more advantageous than other types of sensors. The active sensing materials used in these types of sensors can be any material that acts as a catalyst for the oxidation or reduction of particular analyte or set of analytes. Among various kinds of sensing materials, deoxyribonucleic acid (DNA), carbon nanotubes (CNTs) and nanoparticles have received considerable attraction in recent years. DNA is one of the classes of natural polymers, which can interact with CNTs and nanoparticles to form new types of composite materials. These composite materials have also been used as sensing materials for sensor applications. They have advantages in characteristics such as extraordinary low weight and multifunctional properties. In this article, advantages of DNA incorporated in CNT and nanoparticle hybrids for electrochemical sensors and biosensors are presented in detail, along with some key results noted from the literature.

  10. Electrochemical functionalization of polypyrrole through amine oxidation of poly(amidoamine) dendrimers: Application to DNA biosensor.

    PubMed

    Miodek, Anna; Mejri-Omrani, Nawel; Khoder, Rabih; Korri-Youssoufi, Hafsa

    2016-07-01

    Electrochemical patterning method has been developed to fabricate composite based on polypyrrole (PPy) film and poly(amidoamine) dendrimers of fourth generation (PAMAM G4). PPy layer was generated using electrochemical polymerization of pyrrole on a gold electrode. PPy film was then modified with PAMAM G4 using amines electro-oxidation method. Covalent bonding of PAMAM G4 and the formation of PPy-PAMAM composite was characterized using Fourier Transform Infrared Spectroscopy (FT-IR) and X-ray Photoelectron Spectroscopy (XPS). Ferrocenyl groups were then attached to such surface as a redox marker. Electrochemical properties of the modified nanomaterial (PPy-PAMAM-Fc) were studied using both amperometric and impedimetric methods to demonstrate the efficiency of electron transfer through the modified PPy layer. The obtained electrical and electrochemical properties were compared to a composite where PPy bearing carboxylic acid functions was chemically modified with PAMAM G4 by covalent attachment through formation of amid bond (PPy-CONH-PAMAM). The above mentioned studies showed that electrochemical patterning does not disturb the electronic properties of PPy. The effect of the number of functional groups introduced by the electrochemical patterning was demonstrated through the association of various compounds (ethylenediamine, PAMAM G2 and PAMAM G6). We demonstrated that such compounds could be applied in the biosensors technology. The modified PPy-PAMAM-Fc was evaluated as a platform for DNA sensing. High performance in the DNA detection by variation of the electrochemical signal of ferrocene was obtained with detection limit of 0.4 fM. Furthermore, such approach of electrochemical patterning by oxidation of amines could be applied for chemical modification of PPy and open a new way in various biosensing application involving functionalized PPy. PMID:27154698

  11. A novel electrochemical DNA biosensor construction based on layered CuS-graphene composite and Au nanoparticles.

    PubMed

    Xu, Chun-Xuan; Zhai, Qiu-Ge; Liu, Yu-Jie; Huang, Ke-Jing; Lu, Lu; Li, Ke-Xin

    2014-11-01

    A novel CuS-graphene (CuS-Gr) composite was synthesized to achieve excellent electrochemical properties for application as a DNA electrochemical biosensor. CuS-Gr composite was prepared by a hydrothermal method, in which two-dimensional graphene served as a two-dimensional conductive skeleton to support CuS nanoparticles. A sensitive electrochemical DNA biosensor was fabricated by immobilizing single-stranded DNA (ss-DNA) labeled at the 5' end using 6-mercapto-1-hexane (HS-ssDNA) on the surface of Au nanoparticles (AuNPs) to form ssDNA-S-AuNPs/CuS-Gr, and hybridization sensing was done in phosphate buffer. Cyclic voltammetry and electrochemical impedance spectroscopy were performed for the characterization of the modified electrodes. Differential pulse voltammetry was applied to monitor the DNA hybridization using an [Fe(CN)6](3-/4-) solution as a probe. Under optimum conditions, the biosensor developed exhibited a good linear relationship between the current and the logarithm of the target DNA concentration ranging from 0.001 to 1 nM, with a low detection limit of 0.1 pM (3σ/S). The biosensor exhibited high selectivity to differentiate one-base-mismatched DNA and three-base-mismatched DNA. The results indicated that the sensing platform based on CuS-Gr provides a stable and conductive interface for electrochemical detection of DNA hybridization, and could easily be extended to the detection of other nucleic acids. PMID:24894519

  12. Nanostructured electrochemical DNA biosensors for detection of the effect of berberine on DNA from cancer cells.

    PubMed

    Ovádeková, Renáta; Jantová, Sona; Letasiová, Silvia; Stepánek, Ivan; Labuda, Ján

    2006-12-01

    Multi walled carbon nanotubes (MWNT) in dimethylformamide (DMF) or aqueous sodium dodecyl sulfate (SDS) solution, colloidal gold nanoparticles (GNP) in phosphate buffer solution (PBS), and a GNP-MWNT mixture in aqueous SDS solution have been investigated for chemical modification of a screen-printed carbon electrode used as the signal transducer of a dsDNA-based biosensor. Differential pulse voltammetry of the DNA redox marker Co[(phen)3]3+ and the guanine moiety anodic oxidation and cyclic voltammetry with K3[Fe(CN)6] as indicator revealed substantial enhancement of the response of the biosensor, particularly when MWNT in SDS solution was used. The biosensor was used in testing of berberine, an isoquinoline plant alkaloid with significant antimicrobial and anticancer activity. Berberine had a very strong, concentration-dependent, effect on the structural stability of DNA from the human cancer cells (U937 cells) whereas non-cancer cells were changed only when berberine concentrations were relatively high 75 and 50 microg mL(-1). PMID:17053918

  13. Ultraspecific electrochemical DNA biosensor by coupling spontaneous cascade DNA branch migration and dual-signaling sensing strategy.

    PubMed

    Wang, Ting; Zhou, Lili; Bai, Shulian; Zhang, Zhang; Li, Junlong; Jing, Xiaoying; Xie, Guoming

    2016-04-15

    Using spontaneous cascade DNA branch migration and dual-signaling sensing strategy, we developed a novel universal electrochemical biosensor for the highly specific and sensitive detection of nucleic acids. A target strand (Ts) competitively hybridized with a ferrocene (Fc)-labeled signal probe (Fc-S1), which was blocked by a protector strand (Ps), after strand displacement to form the Ts/Fc-S1 duplex. A methylene blue (MB)-modified signal probe (MB-S2) was immobilized on the Au electrode surface by hybridizing with a thiolated capture probe (Cp). Then, the obtained reactants (Ts/Fc-S1 and MB-S2/Cp) suffered spontaneous DNA branch migration and produced two hybridization products (Fc-S1/Cp and MB-S2/Ts). These reactions led to the dissociation of MB molecules and the collection of Fc molecules. The detection mechanism of this DNA biosensor involved distance variation between the redox tags and the Au electrode, which was associated with target-induced cascade DNA branch migration. Moreover, we rationally designed the cascade DNA branch migration to occur spontaneously with ΔG° ≈ 0, at which slight thermodynamic changes caused by base mismatch exerted a disproportionately large effect on the hybridization yield. This "signal-on/off" sensing system exhibited a remarkable analytical performance and an ultrahigh discrimination capability even against a single-base mismatch. The maximum discrimination factor (DF) of base mutations or alterations can reach 17.9. Therefore, our electrochemical biosensor might hold a great potential for further applications in biomedical research and early clinical diagnosis. PMID:26657589

  14. Electrochemical DNA biosensor for detection of porcine oligonucleotides using ruthenium(II) complex as intercalator label redox

    NASA Astrophysics Data System (ADS)

    Halid, Nurul Izni Abdullah; Hasbullah, Siti Aishah; Ahmad, Haslina; Heng, Lee Yook; Karim, Nurul Huda Abd; Harun, Siti Norain

    2014-09-01

    A DNA biosensor detection of oligonucleotides via the interactions of porcine DNA with redox active complex based on the electrochemical transduction is described. A ruthenium(II) complex, [Ru(bpy)2(PIP)]2+, (bpy = 2,2'bipyridine, PIP = 2-phenylimidazo[4,5-f[[1,10-phenanthroline]) as DNA label has been synthesized and characterized by 1H NMR and mass spectra. The study was carried out by covalent bonding immobilization of porcine aminated DNA probes sequences on screen printed electrode (SPE) modified with succinimide-acrylic microspheres and [Ru(bpy)2(PIP)]2+ was used as electrochemical redox intercalator label to detect DNA hybridization event. Electrochemical detection was performed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) over the potential range where the ruthenium (II) complex was active. The results indicate that the interaction of [Ru(bpy)2(PIP)]2+ with hybridization complementary DNA has higher response compared to single-stranded and mismatch complementary DNA.

  15. Development of an electrochemical biosensor methods based on acrylic microsphere for the determination of Arowana DNA hybridization

    NASA Astrophysics Data System (ADS)

    Rahman, Mahbubur; Heng, Lee Yook; Futra, Dedi; Chiang, Chew Poh

    2015-09-01

    An electrochemical method of Arowana DNA determination based of N-acrylosuccinimide (NAS) modified acrylic microsphere was fabricated. Hydrophobic succinimide functional group containing poly(n-butylacrylate-N-acryloxysuccinimide) microspheres were synthesized with a simple one-step photopolymerization pocedure. Aminated DNA probe was covalently bonded to the succinimde functional group of the acrylic microspheres. The hybridization of the immobilized DNA probe with the complementary DNA was determined by the differential pulse voltametry using anthraquninone-2-sulfonic acid monohydrate sodium salt (AQMS) as the electroactive hybridization label. The influences of many factors such as duration of DNA probe immobilization and hybridization, operational temperature and non-complementary DNA on the biosensor performance were evaluated. Under optimized conditions, the DNA microbiosensor demonstrated a wide linear response range to target DNA is 1.0 × 10-16 and 1.0 × 10-8 M with a lower limit of detection (LOD) of 9.46 × 10-17 M (R2 = 0.99) were calculated. This biosensor had improved the overall analytical performance of the resultant DNA microbiosensor when compared with other reported DNA biosensors using other nano-materials for membranes and microspheres as DNA immobilization matrices.

  16. Electrochemical biosensors and nanobiosensors.

    PubMed

    Hammond, Jules L; Formisano, Nello; Estrela, Pedro; Carrara, Sandro; Tkac, Jan

    2016-06-30

    Electrochemical techniques have great promise for low-cost miniaturised easy-to-use portable devices for a wide range of applications-in particular, medical diagnosis and environmental monitoring. Different techniques can be used for biosensing, with amperometric devices taking the central role due to their widespread application in glucose monitoring. In fact, glucose biosensing takes an approximately 70% share of the biosensor market due to the need for diabetic patients to monitor their sugar levels several times a day, making it an appealing commercial market.In this review, we present the basic principles of electrochemical biosensor devices. A description of the different generations of glucose sensors is used to describe in some detail the operation of amperometric sensors and how the introduction of mediators can enhance the performance of the sensors. Electrochemical impedance spectroscopy is a technique being increasingly used in devices due to its ability to detect variations in resistance and capacitance upon binding events. Novel advances in electrochemical sensors, due to the use of nanomaterials such as carbon nanotubes and graphene, are presented as well as future directions that the field is taking. PMID:27365037

  17. Electrochemical biosensors and nanobiosensors

    PubMed Central

    Hammond, Jules L.; Formisano, Nello; Carrara, Sandro; Tkac, Jan

    2016-01-01

    Electrochemical techniques have great promise for low-cost miniaturised easy-to-use portable devices for a wide range of applications–in particular, medical diagnosis and environmental monitoring. Different techniques can be used for biosensing, with amperometric devices taking the central role due to their widespread application in glucose monitoring. In fact, glucose biosensing takes an approximately 70% share of the biosensor market due to the need for diabetic patients to monitor their sugar levels several times a day, making it an appealing commercial market. In this review, we present the basic principles of electrochemical biosensor devices. A description of the different generations of glucose sensors is used to describe in some detail the operation of amperometric sensors and how the introduction of mediators can enhance the performance of the sensors. Electrochemical impedance spectroscopy is a technique being increasingly used in devices due to its ability to detect variations in resistance and capacitance upon binding events. Novel advances in electrochemical sensors, due to the use of nanomaterials such as carbon nanotubes and graphene, are presented as well as future directions that the field is taking. PMID:27365037

  18. Conducting polymer based electrochemical biosensors.

    PubMed

    Aydemir, Nihan; Malmström, Jenny; Travas-Sejdic, Jadranka

    2016-03-28

    Conducting polymer (CP)-based electrochemical biosensors have gained great attention as such biosensor platforms are easy and cost-effective to fabricate, and provide a direct electrical readout for the presence of biological analytes with high sensitivity and selectivity. CP materials themselves are both sensing elements and transducers of the biological recognition event at the same time, simplifying sensor designs. This review summarizes the advances in electrochemical biosensors based on CPs. Recognition probe immobilisation techniques, transduction mechanisms and detection of various target biomolecules have been discussed in detail. Efforts to miniaturize CP-based electrochemical biosensors and fabrication of sensor arrays are also briefly reviewed. PMID:26948182

  19. An electrochemical DNA biosensor based on nitrogen-doped graphene/Au nanoparticles for human multidrug resistance gene detection.

    PubMed

    Chen, Mei; Hou, Changjun; Huo, Danqun; Bao, Jing; Fa, Huanbao; Shen, Caihong

    2016-11-15

    Multidrug resistance (MDR) has become a major obstacle to the adequate treatment of cancer patients; thus, there is an urgent need for exploring new strategies for early diagnosis of MDR in clinic. Here, we report a novel electrochemical biosensor based on nitrogen-doped graphene nanosheets functionalized with Au nanoparticles (N-G/Au) for sensitive and selective DNA detection. The highly conductive nanocomposite layer was characterized by using scanning and transmission electron microscopy, X-ray photoelectron spectroscopy, and cyclic voltammetry. DNA with thiol groups at the 5' end was immobilized on the N-G/Au surface via the strong Au-S bond. Differential pulse voltammetry was applied to monitor the target DNA hybridization event using methylene blue as an electrochemical indicator. Under optimal conditions, the biosensor could detect target DNA down to 3.12×10(-15)M with a linear range from 1.0×10(-14) to 1.0×10(-7)M, showing high sensitivity. Further, the sensing strategy was successfully used for detecting MDR1 DNA in real clinical samples. These results will aid in developing a new portable detection system for MDR that will allow effective diagnosis in the early stages of related cancer. PMID:27258172

  20. A highly selective and sensitive electrochemical CS-MWCNTs/Au-NPs composite DNA biosensor for Staphylococcus aureus gene sequence detection.

    PubMed

    Sun, Yange; He, Xingxing; Ji, Jian; Jia, Min; Wang, Zhouping; Sun, Xiulan

    2015-08-15

    This paper presents a new electrochemical DNA biosensor constructed using a substrate electrode composed of a novel nanocomposite material prepared using gold nanoparticles (Au-NPs) and multiwalled carbon nanotubes (MWCNTs) and further modified with an Au electrode (AuE), which was used as the substrate electrode. A single-stranded DNA (ssDNA) probe was immobilized on the Au-NPs/CS-MWCNTs/AuE electrode by means of facile gold-thiol affinity, which resulted in hybridization with the target ssDNA sequence. Hybridization reactions were assessed by using the reduction peak current of methylene blue (MB) as an electrochemical indicator. The advantages of the nanomaterials were found to include high surface area, favorable electronic properties, and strong electrocatalytic activity. The amount of ssDNA adsorbed on the electrode surface was increased and the electrochemical response of MB accelerated. The differential pulse voltammetric responses of MB were in line with the specific target ssDNA sequence within the concentration range 1.0×10(-15)-1.0×10(-8)M with the detection limit 3.3×10(-16)M (3σ). In the colony forming unit (CFU) we were able to detect 10CFU mL(-1)of Staphylococcus aureus in the tap water, achieving good discrimination ability between one- and three-base mismatched ssDNA sequences. The polymerase chain reaction (PCR) amplification products of S. aureus nuc gene sequence were also detected with satisfactory results. PMID:25966418

  1. Electrochemical DNA biosensor for detection of porcine oligonucleotides using ruthenium(II) complex as intercalator label redox

    SciTech Connect

    Halid, Nurul Izni Abdullah; Hasbullah, Siti Aishah; Heng, Lee Yook; Karim, Nurul Huda Abd; Ahmad, Haslina; Harun, Siti Norain

    2014-09-03

    A DNA biosensor detection of oligonucleotides via the interactions of porcine DNA with redox active complex based on the electrochemical transduction is described. A ruthenium(II) complex, [Ru(bpy){sub 2}(PIP)]{sup 2+}, (bpy = 2,2′bipyridine, PIP = 2-phenylimidazo[4,5-f[[1,10-phenanthroline]) as DNA label has been synthesized and characterized by 1H NMR and mass spectra. The study was carried out by covalent bonding immobilization of porcine aminated DNA probes sequences on screen printed electrode (SPE) modified with succinimide-acrylic microspheres and [Ru(bpy){sub 2}(PIP)]{sup 2+} was used as electrochemical redox intercalator label to detect DNA hybridization event. Electrochemical detection was performed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) over the potential range where the ruthenium (II) complex was active. The results indicate that the interaction of [Ru(bpy){sub 2}(PIP)]{sup 2+} with hybridization complementary DNA has higher response compared to single-stranded and mismatch complementary DNA.

  2. A Sensitive and Selective Label-Free Electrochemical DNA Biosensor for the Detection of Specific Dengue Virus Serotype 3 Sequences

    PubMed Central

    Oliveira, Natália; Souza, Elaine; Ferreira, Danielly; Zanforlin, Deborah; Bezerra, Wessulla; Borba, Maria Amélia; Arruda, Mariana; Lopes, Kennya; Nascimento, Gustavo; Martins, Danyelly; Cordeiro, Marli; Lima-Filho, José

    2015-01-01

    Dengue fever is the most prevalent vector-borne disease in the world, with nearly 100 million people infected every year. Early diagnosis and identification of the pathogen are crucial steps for the treatment and for prevention of the disease, mainly in areas where the co-circulation of different serotypes is common, increasing the outcome of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Due to the lack of fast and inexpensive methods available for the identification of dengue serotypes, herein we report the development of an electrochemical DNA biosensor for the detection of sequences of dengue virus serotype 3 (DENV-3). DENV-3 probe was designed using bioinformatics software and differential pulse voltammetry (DPV) was used for electrochemical analysis. The results showed that a 22-m sequence was the best DNA probe for the identification of DENV-3. The optimum concentration of the DNA probe immobilized onto the electrode surface is 500 nM and a low detection limit of the system (3.09 nM). Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions. Therefore, the application of DNA biosensors for diagnostics at the molecular level may contribute to future advances in the implementation of specific, effective and rapid detection methods for the diagnosis dengue viruses. PMID:26140346

  3. A Sensitive and Selective Label-Free Electrochemical DNA Biosensor for the Detection of Specific Dengue Virus Serotype 3 Sequences.

    PubMed

    Oliveira, Natália; Souza, Elaine; Ferreira, Danielly; Zanforlin, Deborah; Bezerra, Wessulla; Borba, Maria Amélia; Arruda, Mariana; Lopes, Kennya; Nascimento, Gustavo; Martins, Danyelly; Cordeiro, Marli; Lima-Filho, José

    2015-01-01

    Dengue fever is the most prevalent vector-borne disease in the world, with nearly 100 million people infected every year. Early diagnosis and identification of the pathogen are crucial steps for the treatment and for prevention of the disease, mainly in areas where the co-circulation of different serotypes is common, increasing the outcome of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Due to the lack of fast and inexpensive methods available for the identification of dengue serotypes, herein we report the development of an electrochemical DNA biosensor for the detection of sequences of dengue virus serotype 3 (DENV-3). DENV-3 probe was designed using bioinformatics software and differential pulse voltammetry (DPV) was used for electrochemical analysis. The results showed that a 22-m sequence was the best DNA probe for the identification of DENV-3. The optimum concentration of the DNA probe immobilized onto the electrode surface is 500 nM and a low detection limit of the system (3.09 nM). Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions. Therefore, the application of DNA biosensors for diagnostics at the molecular level may contribute to future advances in the implementation of specific, effective and rapid detection methods for the diagnosis dengue viruses. PMID:26140346

  4. Biosensors for DNA sequence detection

    NASA Technical Reports Server (NTRS)

    Vercoutere, Wenonah; Akeson, Mark

    2002-01-01

    DNA biosensors are being developed as alternatives to conventional DNA microarrays. These devices couple signal transduction directly to sequence recognition. Some of the most sensitive and functional technologies use fibre optics or electrochemical sensors in combination with DNA hybridization. In a shift from sequence recognition by hybridization, two emerging single-molecule techniques read sequence composition using zero-mode waveguides or electrical impedance in nanoscale pores.

  5. Nanomaterial-Based Electrochemical Biosensors and Bioassays

    SciTech Connect

    Liu, Guodong; Mao, Xun; Gurung, Anant; Baloda, Meenu; Lin, Yuehe; He, Yuqing

    2010-08-31

    This book chapter summarizes the recent advance in nanomaterials for electrochemical biosensors and bioassays. Biofunctionalization of nanomaterials for biosensors fabrication and their biomedical applications are discussed.

  6. A label-free electrochemical DNA biosensor based on covalent immobilization of salmonella DNA sequences on the nanoporous glassy carbon electrode.

    PubMed

    Amouzadeh Tabrizi, Mahmoud; Shamsipur, Mojtaba

    2015-07-15

    Herein, an easy and cost-effective approach to the immobilization of probe was performed. The amino modified salmonella ssDNA probe sequence was covalently linked with carboxylic group on the surface of nanoporous glassy carbon electrode to prepare the DNA biosensor. The differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) techniques were used for the determination of salmonella DNA in the concentration ranges of 10-400pM and 1-400pM with limits of detection of 2.1pM and 0.15pM, respectively. PMID:25710894

  7. Electrochemical study of quinone redox cycling: A novel application of DNA-based biosensors for monitoring biochemical reactions.

    PubMed

    Ensafi, Ali A; Jamei, Hamid Reza; Heydari-Bafrooei, Esmaeil; Rezaei, B

    2016-10-01

    This paper presents the results of an experimental investigation of voltammetric and impedimetric DNA-based biosensors for monitoring biological and chemical redox cycling reactions involving free radical intermediates. The concept is based on associating the amounts of radicals generated with the electrochemical signals produced, using differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). For this purpose, a pencil graphite electrode (PGE) modified with multiwall carbon nanotubes and poly-diallydimethlammonium chloride decorated with double stranded fish sperm DNA was prepared to detect DNA damage induced by the radicals generated from a redox cycling quinone (i.e., menadione (MD; 2-methyl-1,4-naphthoquinone)). Menadione was employed as a model compound to study the redox cycling of quinones. A direct relationship was found between free radical production and DNA damage. The relationship between MD-induced DNA damage and free radical generation was investigated in an attempt to identify the possible mechanism(s) involved in the action of MD. Results showed that DPV and EIS were appropriate, simple and inexpensive techniques for the quantitative and qualitative comparisons of different reducing reagents. These techniques may be recommended for monitoring DNA damages and investigating the mechanisms involved in the production of redox cycling compounds. PMID:27179196

  8. Overview of Electrochemical DNA Biosensors: New Approaches to Detect the Expression of Life

    PubMed Central

    Cagnin, Stefano; Caraballo, Marcelo; Guiducci, Carlotta; Martini, Paolo; Ross, Marty; SantaAna, Mark; Danley, David; West, Todd; Lanfranchi, Gerolamo

    2009-01-01

    DNA microarrays are an important tool with a variety of applications in gene expression studies, genotyping, pharmacogenomics, pathogen classification, drug discovery, sequencing and molecular diagnostics. They are having a strong impact in medical diagnostics for cancer, toxicology and infectious disease applications. A series of papers have been published describing DNA biochips as alternative to conventional microarray platforms to facilitate and ameliorate the signal readout. In this review, we will consider the different methods proposed for biochip construction, focusing on electrochemical detection of DNA. We also introduce a novel single-stranded DNA platform performing high-throughput SNP detection and gene expression profiling. PMID:22574066

  9. Fabrication of an electrochemical DNA-based biosensor for Bacillus cereus detection in milk and infant formula.

    PubMed

    Izadi, Zahra; Sheikh-Zeinoddin, Mahmoud; Ensafi, Ali A; Soleimanian-Zad, Sabihe

    2016-06-15

    This paper describes fabrication of a DNA-based Au-nanoparticle modified pencil graphite electrode (PGE) biosensor for detection of Bacillus cereus, causative agent of two types of food-borne disease, i.e., emetic and diarrheal syndrome. The sensing element of the biosensor was comprised of gold nanoparticles (GNPs) self-assembled with single-stranded DNA (ssDNA) of nheA gene immobilized with thiol linker on the GNPs modified PGE. The size, shape and dispersion of the GNPs were characterized by field emission scanning electron microscope (FESEM). Detection of B. cereus was carried out based on an increase in the charge transfer resistance (Rct) of the biosensor due to hybridization of the ss-DNA with target DNA. An Atomic force microscope (AFM) was used to confirm the hybridization. The biosensor sensitivity in pure cultures of B. cereus was found to be 10(0) colony forming units per milliliter (CFU/mL) with a detection limit of 9.4 × 10(-12) mol L(-1). The biosensor could distinguish complementary from mismatch DNA sequence. The proposed biosensor exhibited a rapid detection, low cost, high sensitivity to bacterial contamination and could exclusively and specifically detect the target DNA sequence of B. cereus from other bacteria that can be found in dairy products. Moreover, the DNA biosensor exhibited high reproducibility and stability, thus it may be used as a suitable biosensor to detect B. cereus and to become a portable system for food quality control. PMID:26896793

  10. A Simple DNA-based Electrochemical Biosensor for Highly Sensitive Detection of Ciprofloxacin Using Disposable Graphene.

    PubMed

    Lim, Syazana A; Ahmed, Minhaz U

    2016-01-01

    In this work we exploited the electrostatic interaction of double stranded DNA (dsDNA) with drug components to construct a simple, but highly sensitive, DNA-electrochemical sensor for detecting ciprofloxacin. The following straightforward three-step procedure was performed to determine ciprofloxacin: (i) dsDNA-layer immobilization on the surface of the working graphene-modified screen-printed carbon electrode; (ii) dsDNA-ciprofloxacin interaction for 2 min; and (iii) electrochemical measurement using square-wave voltammetry. An increased oxidation of the guanine component was observed, at +1.0 V, as a result of the electrostatic interaction of positively charged ciprofloxacin with the negatively charged nucleic acid sugar phosphate. Based on the International Conference on Harmonization Guidelines, a linear relationship between the guanine oxidation peak and ciprofloxacin concentration (0.1 to 100 μM) was obtained with a detection limit of 0.1 μM. Our developed sensor is straightforward to construct and use, requiring no multi-step time-consuming preconditioning of electrodes. It is highly sensitive and selective in the detection of ciprofloxacin, and has the potential to be useful in the future fabrication of rapid and portable on-site food safety analysis devices. PMID:27302591

  11. An electrochemical competitive biosensor for ochratoxin A based on a DNA biotinylated aptamer.

    PubMed

    Bonel, Laura; Vidal, Juan C; Duato, Patricia; Castillo, Juan R

    2011-03-15

    Ochratoxin A (OTA) is one of the most important mycotoxin contaminants of foods, particularly cereals and cereal products, with strict low regulatory levels (of ppb) in many countries worldwide. An electrochemical competitive aptamer-based biosensor for OTA is described. Paramagnetic microparticle beads (MBs) were functionalized with an aptamer specific to OTA, and were allowed to compete with a solution of the mycotoxin conjugated to the enzyme horseradish peroxidase (OTA-HRP) and free OTA. After separation and washing steps helped with magnetic separations, the modified MBs were localized on disposable screen-printed carbon electrodes (SPCEs) under a magnetic field, and the product of the enzymatic reaction with the substrate was detected with differential-pulse voltammetry. In addition to magnetic separation assays, other competitive schemes (direct/indirect aptasensors performed on the SPCEs surface or using gold nanoparticles functionalized with the aptamer) were preliminary tested, optimized and compared. The magnetic aptasensor showed a linear response to OTA in the range 0.78-8.74 ng mL(-1) and a limit of detection of 0.07±0.01 ng mL(-1), and was accurately applied to extracts of certified and spiked wheat samples with an RSD lower than about 8%. PMID:21256729

  12. Ultrasensitive electrochemical biosensor for specific detection of DNA based on molecular beacon mediated circular strand displacement polymerization and hyperbranched rolling circle amplification.

    PubMed

    Li, Xiaolu; Guo, Jing; Zhai, Qian; Xia, Jing; Yi, Gang

    2016-08-31

    Using a cascade signal amplification strategy, an ultrasensitive electrochemical biosensor for specific detection of DNA based on molecular beacon (MB) mediated circular strand displacement polymerization (CSDP) and hyperbranched rolling circle amplification (HRCA) was proposed. The hybridization of MB probe to target DNA resulted in a conformational change of the MB and triggered the CSDP in the presence of bio-primer and Klenow fragment (KF exo(-)), leading to multiple biotin-tagged DNA duplex. Furthermore, the HRCA was implemented to product amounts of double-stranded DNA (ds-DNA) fragments using phi29 DNA polymerase via biotin-streptavidin interaction. After the product of HRCA binded numerous biotinylated detection probes, an ultrasensitive electrochemical readout by further employing the streptavidin-alkaline phosphatase. The proposed biosensor exhibited excellent detection sensitivity and specificity with a log-linear response to target DNA from 0.01 fM to 10 pM as low as 8.9 aM. The proposed method allowed DNA detection with simplicity, rapidness, low cost and high specificity, which might have the potential for application in clinical molecular diagnostics and environmental monitoring. PMID:27506343

  13. A novel electrochemical DNA biosensor based on a modified magnetic bar carbon paste electrode with Fe3O4NPs-reduced graphene oxide/PANHS nanocomposite.

    PubMed

    Jahanbani, Shahriar; Benvidi, Ali

    2016-11-01

    In this study, we have designed a label free DNA biosensor based on a magnetic bar carbon paste electrode (MBCPE) modified with nanomaterial of Fe3O4/reduced graphene oxide (Fe3O4NP-RGO) as a composite and 1- pyrenebutyric acid-N- hydroxysuccinimide ester (PANHS) as a linker for detection of DNA sequences. Probe (BRCA1 5382 insC mutation detection) strands were immobilized on the MBCPE/Fe3O4-RGO/PANHS electrode for the exact incubation time. The characterization of the modified electrode was studied using different techniques such as scanning electron microscopy (SEM), infrared spectroscopy (IR), vibrating sample magnetometer (VSM), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry methods. Some experimental parameters such as immobilization time of probe DNA, time and temperature of hybridization process were investigated. Under the optimum conditions, the immobilization of the probe and its hybridization with the target DNA (Complementary DNA) were tested. This DNA biosensor revealed a good linear relationship between ∆Rct and logarithm of the complementary target DNA concentration ranging from 1.0×10(-18)molL(-1) to 1.0×10(-8)molL(-1) with a correlation coefficient of 0.9935 and a detection limit of 2.8×10(-19)molL(-1). In addition, the mentioned biosensor was satisfactorily applied for discriminating of complementary sequences from non-complementary sequences. The constructed biosensor (MBCPE/Fe3O4-RGO/PANHS/ssDNA) with high sensitivity, selectivity, stability, reproducibility and low cost can be used for detection of BRCA1 5382 insC mutation. PMID:27523989

  14. Electrochemical DNA biosensor based on poly(2,6-pyridinedicarboxylic acid) modified glassy carbon electrode for the determination of anticancer drug gemcitabine.

    PubMed

    Tığ, Gözde Aydoğdu; Zeybek, Bülent; Pekyardımcı, Şule

    2016-07-01

    In this study, a simple methodology was used to develop a new electrochemical DNA biosensor based on poly(2,6-pyridinedicarboxylic acid) (P(PDCA)) modified glassy carbon electrode (GCE). This modified electrode was used to monitor for the electrochemical interaction between the dsDNA and gemcitabine (GEM) for the first time. A decrease in oxidation signals of guanine after the interaction of the dsDNA with the GEM was used as an indicator for the selective determination of the GEM via differential pulse voltammetry (DPV). The guanine oxidation peak currents were linearly proportional to the concentrations of the GEM in the range of 1-30mgL(‒1). Limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.276mgL(‒1) and 0.922mgL(‒1), respectively. The reproducibility, repeatability, and applicability of the analysis to pharmaceutical dosage forms and human serum samples were also examined. In addition to DPV method, UV-vis and viscosity measurements were utilized to propose the interaction mechanism between the GEM and the dsDNA. The novel DNA biosensor could serve for sensitive, accurate and rapid determination of the GEM. PMID:27154680

  15. Recent advances in electrochemical biosensors based on graphene two-dimensional nanomaterials.

    PubMed

    Song, Yang; Luo, Yanan; Zhu, Chengzhou; Li, He; Du, Dan; Lin, Yuehe

    2016-02-15

    Graphene as a star among two-dimensional nanomaterials has attracted tremendous research interest in the field of electrochemistry due to their intrinsic properties, including the electronic, optical, and mechanical properties associated with their planar structure. The marriage of graphene and electrochemical biosensors has created many ingenious biosensing strategies for applications in the areas of clinical diagnosis and food safety. This review provides a comprehensive overview of the recent advances in the development of graphene based electrochemical biosensors. Special attention is paid to graphene-based enzyme biosensors, immunosensors, and DNA biosensors. Future perspectives on high-performance graphene-based electrochemical biosensors are also discussed. PMID:26187396

  16. Real-time, multiplexed electrochemical DNA detection using an active complementary metal-oxide-semiconductor biosensor array with integrated sensor electronics.

    PubMed

    Levine, Peter M; Gong, Ping; Levicky, Rastislav; Shepard, Kenneth L

    2009-03-15

    Optical biosensing based on fluorescence detection has arguably become the standard technique for quantifying extents of hybridization between surface-immobilized probes and fluorophore-labeled analyte targets in DNA microarrays. However, electrochemical detection techniques are emerging which could eliminate the need for physically bulky optical instrumentation, enabling the design of portable devices for point-of-care applications. Unlike fluorescence detection, which can function well using a passive substrate (one without integrated electronics), multiplexed electrochemical detection requires an electronically active substrate to analyze each array site and benefits from the addition of integrated electronic instrumentation to further reduce platform size and eliminate the electromagnetic interference that can result from bringing non-amplified signals off chip. We report on an active electrochemical biosensor array, constructed with a standard complementary metal-oxide-semiconductor (CMOS) technology, to perform quantitative DNA hybridization detection on chip using targets conjugated with ferrocene redox labels. A 4 x 4 array of gold working electrodes and integrated potentiostat electronics, consisting of control amplifiers and current-input analog-to-digital converters, on a custom-designed 5 mm x 3 mm CMOS chip drive redox reactions using cyclic voltammetry, sense DNA binding, and transmit digital data off chip for analysis. We demonstrate multiplexed and specific detection of DNA targets as well as real-time monitoring of hybridization, a task that is difficult, if not impossible, with traditional fluorescence-based microarrays. PMID:19054661

  17. DNA biosensors that reason.

    PubMed

    Sainz de Murieta, Iñaki; Rodríguez-Patón, Alfonso

    2012-08-01

    Despite the many designs of devices operating with the DNA strand displacement, surprisingly none is explicitly devoted to the implementation of logical deductions. The present article introduces a new model of biosensor device that uses nucleic acid strands to encode simple rules such as "IF DNA_strand(1) is present THEN disease(A)" or "IF DNA_strand(1) AND DNA_strand(2) are present THEN disease(B)". Taking advantage of the strand displacement operation, our model makes these simple rules interact with input signals (either DNA or any type of RNA) to generate an output signal (in the form of nucleotide strands). This output signal represents a diagnosis, which either can be measured using FRET techniques, cascaded as the input of another logical deduction with different rules, or even be a drug that is administered in response to a set of symptoms. The encoding introduces an implicit error cancellation mechanism, which increases the system scalability enabling longer inference cascades with a bounded and controllable signal-noise relation. It also allows the same rule to be used in forward inference or backward inference, providing the option of validly outputting negated propositions (e.g. "diagnosis A excluded"). The models presented in this paper can be used to implement smart logical DNA devices that perform genetic diagnosis in vitro. PMID:22406690

  18. DNA nanotechnology-enabled biosensors.

    PubMed

    Chao, Jie; Zhu, Dan; Zhang, Yinan; Wang, Lianhui; Fan, Chunhai

    2016-02-15

    Biosensors employ biological molecules to recognize the target and utilize output elements which can translate the biorecognition event into electrical, optical or mass-sensitive signals to determine the quantities of the target. DNA-based biosensors, as a sub-field to biosensor, utilize DNA strands with short oligonucleotides as probes for target recognition. Although DNA-based biosensors have offered a promising alternative for fast, simple and cheap detection of target molecules, there still exist key challenges including poor stability and reproducibility that hinder their competition with the current gold standard for DNA assays. By exploiting the self-recognition properties of DNA molecules, researchers have dedicated to make versatile DNA nanostructures in a highly rigid, controllable and functionalized manner, which offers unprecedented opportunities for developing DNA-based biosensors. In this review, we will briefly introduce the recent advances on design and fabrication of static and dynamic DNA nanostructures, and summarize their applications for fabrication and functionalization of DNA-based biosensors. PMID:26212206

  19. A microfluidic electrochemical biosensor based on multiwall carbon nanotube/ferrocene for genomic DNA detection of Mycobacterium tuberculosis in clinical isolates.

    PubMed

    Zribi, B; Roy, E; Pallandre, A; Chebil, S; Koubaa, M; Mejri, N; Magdinier Gomez, H; Sola, C; Korri-Youssoufi, H; Haghiri-Gosnet, A-M

    2016-01-01

    Herein we present a microfluidic-multiplexed platform that integrates electrochemical sensors based on carbon nanotubes associated with ferrocene as redox marker (carbon nanotube (CNT)/ferrocene) for direct detection of pathogenic viral DNA from Hepatitis C and genomic DNA from Mycobacterium tuberculosis in clinical isolates. By operating the fluidic device under high flow (150 μl/min), the formation of a very thin depletion layer at the sensor surface (δS = 230 nm) enhances the capture rate up to one DNA strand per second. By comparison, this capture rate is only 0.02 molecule/s in a static regime without flow. This fluidic protocol allows thus enhancing the limit of detection of the electrochemical biosensor from picomolar in bulk solution to femtomolar with a large dynamic range from 0.1 fM to 1 pM. Kinetics analysis also demonstrates an enhancement of the rate constant of electron transfer (kS) of the electrochemical process from 1 s(-1) up to 6 s(-1) thanks to the geometry of the miniaturized fluidic electrochemical cell. This microfluidic device working under high flow allows selective direct detection of a Mycobacterium tuberculosis (H37Rv) rpoB allele from clinical isolate extracted DNA. We envision that a microfluidic approach under high flow associated with a multiwall CNT/ferrocene sensor could find useful applications as the point-of-care for multi-target diagnostics of biomarkers in real samples. PMID:26865908

  20. Graphene Based Electrochemical Sensors and Biosensors: A Review

    SciTech Connect

    Shao, Yuyan; Wang, Jun; Wu, Hong; Liu, Jun; Aksay, Ilhan A.; Lin, Yuehe

    2010-05-01

    Graphene, emerging as a true 2-dimensional material, has received increasing attention due to its unique physicochemical properties (high surface area, excellent conductivity, high mechanical strength, and ease of functionalization and mass production). This article selectively reviews recent advances in graphene-based electrochemical sensors and biosensors. In particular, graphene for direct electrochemistry of enzyme, its electrocatalytic activity toward small biomolecules (hydrogen peroxide, NADH, dopamine, etc.), and graphene-based enzyme biosensors have been summarized in more detail; Graphene-based DNA sensing and environmental analysis have been discussed. Future perspectives in this rapidly developing field are also discussed.

  1. An electrochemical biosensor based on DNA tetrahedron/graphene composite film for highly sensitive detection of NADH.

    PubMed

    Li, Zonglin; Su, Wenqiong; Liu, Shuopeng; Ding, Xianting

    2015-07-15

    Dihydronicotinamide adenine dinucleotide (NADH) is a major biomarker correlated with lethal diseases such as cancers and bacterial infection. Herein, we report a graphene-DNA tetrahedron-gold nanoparticle modified gold disk electrode for highly sensitive NADH detection. By assembling the DNA tetrahedron/graphene composite film on the gold disk electrode surface which prior harnessed electrochemical deposition of gold nanoparticles to enhance the effective surface area, the oxidation potential of NADH was substantially decreased to 0.28V (vs. Ag/AgCl) and surface fouling effects were successfully eliminated. Furthermore, the lower detection limit of NADH by the presented platform was reduced down to 1fM, with an upper limit of 10pM. Both the regeneration and selectivity of composite film-modified electrode are investigated and proved to be robust. The novel sensor developed here could serve as a highly sensitive probe for NADH detection, which would further benefit the field of NADH related disease diagnostics. PMID:25770460

  2. Assessment of genotoxicity of catecholics using impedimetric DNA-biosensor.

    PubMed

    Ensafi, Ali A; Amini, Maryam; Rezaei, B

    2014-03-15

    The potential toxicity of catecholics is a big concern, because the catechol-derived semiquinone radical after the oxidation of catechol (CA) can donate an H-atom to generate quinone, and during this process a superoxide anion radical may be produced. Considering the fact that catecholics are highly consumed in our daily life and some drugs also contain one or more CA moieties, we speculate that CA's toxicity might not be insurmountable. Therefore, finding approaches to investigate catecholics potential toxicity is of great significance. Here in, an electrochemical protocol for direct monitoring of genotoxicity of catecholics is described. CA encapsulated on MWCNTs (CA@MWCNT) through continuous cyclic voltammetric on the surface of pencil graphite electrode (PGE). Subsequently, a DNA functionalized biosensor (DNA/CA@MWCNT/PGE) was prepared and characterized for the detection and the investigation of DNA damage induced by radicals generated from catecholics. The change in the charge transfer resistance (Rct) after the incubation of the DNA biosensor in the damaging solution for a certain time was used as an indicator for DNA damage. Incubation of DNA-modified electrode with CA solution containing Cu(II), Cr(VI) and Fe(III) has been shown to result in oxidative damage to the DNA and change in the electrochemical properties. It was found that the presence of Cu(II), Cr(VI) and Fe(III) in solution caused damage to DNA. The inhibitory effect of glutathione and plumbagin on the CA-mediated DNA damage has also been investigated using the biosensor. The minimum concentration of the metal ions for CA induced DNA damage was investigated. Recognition of suitable matrixes for CA-mediated DNA damage can be assessed using proposed DNA biosensor. Such direct monitoring of the DNA damage holds great promise for designing new biosensors with modification of the biosensor with different damaging agents. PMID:24121207

  3. DNA nanostructures based biosensor for the determination of aromatic compounds.

    PubMed

    Gayathri, S Baby; Kamaraj, P; Arthanareeswari, M; Devikala, S

    2015-10-15

    Graphite electrode was modified using multi-walled carbon nanotubes (MWCNT), chitosan (CS), glutaraldehyde (GTA) and DNA nanostructures (nsDNA). DNA nanostructures of 50 nm in size were produced from single DNA template sequence using a simple two step procedure and were confirmed using TEM and AFM analysis. The modified electrode was applied to the electrochemical detection of aromatic compounds using EIS. The modified electrode was characterized using differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). For comparison, electrochemical results derived from single stranded (50 bp length) and double stranded (50 bp length) DNA based biosensors were used. The results indicate that the modified electrode prior to nsDNA immobilization provides a viable platform that effectively promotes electron transfer between nsDNA and the electrode. The mode of binding between the nsDNA and aromatic compounds was investigated using EIS, indicating that the dominant interaction is non-covalent. nsDNA based biosensor was observed to act as an efficient biosensor in selective and sensitive identification of aromatic compounds. PMID:25982727

  4. Nanoparticle bridge DNA biosensor

    NASA Astrophysics Data System (ADS)

    Huang, Hong-Wen

    A new DNA sensing method is demonstrated in which DNA hybridization events lead to the formation of nanoparticle satellites that bridge two electrodes and are detected electrically. The hybridization events are exclusively carried out only on specific locations, the surfaces of C-ssDNA modified 50 nm GNPs. The uniqueness of this work is that only a small number of T-ccDNA molecules (<10) is required to form the nanoparticle satellites, allowing ultra-sensitive DNA sensing. The principle of this new DNA sensing technique has been demonstrated using target DNA and three-base-pair-mismatched DNA in 20nM concentrations. Three single-stranded DNA (ssDNA) system is used in our experiment which includes Capture-ssDNA (C-ssDNA), Target-ssDNA (T-ssDNA) and Probe-ssDNA (P-ssDNA). Both C-ssDNA and P-ssDNA are modified by a thiol group and can hybridize with different portions of T-ssDNA. T-ssDNA requires no modification in three ssDNA system, which is beneficial in many applications. C-ssDNA modified 50nm gold nanoparticle (C-50au) and P-ssDNA modified 30nm gold nanoparticle (P-30au) are prepared through the reaction of thiol-gold chemical bonding between thiolated ssDNA and gold nanoparticle (GNP) (C-ssDNA with 50nm GNP, P-ssDNA with 30nm GNP). We controllably place the C-50au only on the SiO2 band surface (˜ 90nm width) between two gold electrodes (source and drain electrodes) by forming positively- and negatively-charged self-assembled monolayers (SAMs) on SiO2 and gold surface, respectively. DNA modified GNP is negatively charged due to ionization of phosphate group on DNA back bone. C-50au therefore is negatively charged and can only be attracted toward SiO2 area (repelled by negatively charged gold electrode surface). The amine group of positively-charged SAMs on SiO2 surface is then passivated by converting to non-polar methyl functional group after C-50au placement. P-30au is first hybridized with T-ssDNA in the solution phase (T-P- 30au formed) and is introduced

  5. Synthesis and characterization of nanoparticles for electrochemical biosensor applications

    NASA Astrophysics Data System (ADS)

    Won, Yu-Ho

    Biosensors have been developed for detection, quantification, and monitoring of specific biomolecules or chemical species for environmental, clinical, and industrial fields. Nanoparticles, which can be functionalized by various materials, have attracted research interest in the electrochemical biosensors field due to their versatile physical and chemical properties. Thus, nanoparticles and nanocomposites have been widely investigated as a matrix for the electrochemical biosensors of the detection of various molecules. In this work, nanoparticles, including Fe3O4/silica core/shell nanocomposites, CaCO3-CdSe/ZnS/silica composites, Au nanocrystals, and Cu2O & Cu2O/Au particles, were synthesized and applied for the design of electrochemical biosensors. The goal of this research is to investigate novel nanoparticle-based platforms for the design of highly sensitive and stable biosensors. Biosensors can be categorized into enzyme-based biosensors and enzyme-free biosensors depending on whether or not enzymes are present in the system. Fe3O 4/silica core/shell nanocomposites and CaCO3-CdSe/ZnS/silica composites were used as material platforms to immobilize enzymes and fabricate enzyme-based electrochemical biosensors. On the other hand, Au nanocrystals, Cu2O, and Cu2O/Au particles, which display significant catalytic and electron transfer properties, were investigated in enzyme-free biosensor configurations. In addition, the morphology-dependent biosensing properties of Au nanocrystals, Cu2O, and Cu2O/Au particles were investigated.

  6. Electrochemical Biosensors - Sensor Principles and Architectures

    PubMed Central

    Grieshaber, Dorothee; MacKenzie, Robert; Vörös, Janos; Reimhult, Erik

    2008-01-01

    Quantification of biological or biochemical processes are of utmost importance for medical, biological and biotechnological applications. However, converting the biological information to an easily processed electronic signal is challenging due to the complexity of connecting an electronic device directly to a biological environment. Electrochemical biosensors provide an attractive means to analyze the content of a biological sample due to the direct conversion of a biological event to an electronic signal. Over the past decades several sensing concepts and related devices have been developed. In this review, the most common traditional techniques, such as cyclic voltammetry, chronoamperometry, chronopotentiometry, impedance spectroscopy, and various field-effect transistor based methods are presented along with selected promising novel approaches, such as nanowire or magnetic nanoparticle-based biosensing. Additional measurement techniques, which have been shown useful in combination with electrochemical detection, are also summarized, such as the electrochemical versions of surface plasmon resonance, optical waveguide lightmode spectroscopy, ellipsometry, quartz crystal microbalance, and scanning probe microscopy. The signal transduction and the general performance of electrochemical sensors are often determined by the surface architectures that connect the sensing element to the biological sample at the nanometer scale. The most common surface modification techniques, the various electrochemical transduction mechanisms, and the choice of the recognition receptor molecules all influence the ultimate sensitivity of the sensor. New nanotechnology-based approaches, such as the use of engineered ion-channels in lipid bilayers, the encapsulation of enzymes into vesicles, polymersomes, or polyelectrolyte capsules provide additional possibilities for signal amplification. In particular, this review highlights the importance of the precise control over the delicate

  7. A CMOS Electrochemical Impedance Spectroscopy (EIS) Biosensor Array.

    PubMed

    Manickam, Arun; Chevalier, Aaron; McDermott, Mark; Ellington, Andrew D; Hassibi, Arjang

    2010-12-01

    In this paper, we present a fully integrated biosensor 10 × 10 array in a standard complementary metal-oxide semiconducor process, which takes advantage of electrochemical impedance spectroscopy (EIS). We also show that this system is able to detect various biological analytes, such as DNA and proteins, in real time and without the need for molecular labels. In each pixel of this array, we implement a biocompatible Au electrode transducer and embedded sensor circuitry which takes advantage of the coherent detector to measure the impedance of the associated electrode-electrolyte interface. This chip is capable of concurrently measuring admittance values as small as 10(-8) Ω(-1) within the array with the detection dynamic range of more than 90 dB in the frequency range of 10 Hz-50 MHz. PMID:23850755

  8. Carbon nanomaterial-based electrochemical biosensors: an overview

    NASA Astrophysics Data System (ADS)

    Wang, Zhaoyin; Dai, Zhihui

    2015-04-01

    Carbon materials on the nanoscale exhibit diverse outstanding properties, rendering them extremely suitable for the fabrication of electrochemical biosensors. Over the past two decades, advances in this area have continuously emerged. In this review, we attempt to survey the recent developments of electrochemical biosensors based on six types of carbon nanomaterials (CNs), i.e., graphene, carbon nanotubes, carbon dots, carbon nanofibers, nanodiamonds and buckminsterfullerene. For each material, representative samples are introduced to expound the different roles of the CNs in electrochemical bioanalytical strategies. In addition, remaining challenges and perspectives for future developments are also briefly discussed.

  9. Electrochemical Glucose Biosensor of Platinum Nanospheres Connected by Carbon Nanotubes

    PubMed Central

    Claussen, Jonathan C.; Kim, Sungwon S.; Haque, Aeraj ul; Artiles, Mayra S.; Porterfield, D. Marshall; Fisher, Timothy S.

    2010-01-01

    Background Glucose biosensors comprised of nanomaterials such as carbon nanotubes (CNTs) and metallic nanoparticles offer enhanced electrochemical performance that produces highly sensitive glucose sensing. This article presents a facile biosensor fabrication and biofunctionalization procedure that utilizes CNTs electrochemically decorated with platinum (Pt) nanospheres to sense glucose amperometrically with high sensitivity. Method Carbon nanotubes are grown in situ by microwave plasma chemical vapor deposition (MPCVD) and electro-chemically decorated with Pt nanospheres to form a CNT/Pt nanosphere composite biosensor. Carbon nanotube electrodes are immobilized with fluorescently labeled bovine serum albumin (BSA) and analyzed with fluorescence microscopy to demonstrate their biocompatibility. The enzyme glucose oxidase (GOX) is immobilized onto the CNT/Pt nanosphere biosensor by a simple drop-coat method for amperometric glucose sensing. Results Fluorescence microscopy demonstrates the biofunctionalization capability of the sensor by portraying adsorption of fluorescently labeled BSA unto MPCVD-grown CNT electrodes. The subsequent GOX–CNT/Pt nanosphere biosensor demonstrates a high sensitivity toward H2O2 (7.4 μA/mM/cm2) and glucose (70 μA/mM/cm2), with a glucose detection limit and response time of 380 nM (signal-to-noise ratio = 3) and 8 s (t90%), respectively. The apparent Michaelis–Menten constant (0.64 mM) of the biosensor also reflects the improved sensitivity of the immobilized GOX/nanomaterial complexes. Conclusions The GOX–CNT/Pt nanosphere biosensor outperforms similar CNT, metallic nanoparticle, and more conventional carbon-based biosensors in terms of glucose sensitivity and detection limit. The biosensor fabrication and biofunctionalization scheme can easily be scaled and adapted for microsensors for physiological research applications that require highly sensitive glucose sensing. PMID:20307391

  10. Aptamer-based competitive electrochemical biosensor for brevetoxin-2.

    PubMed

    Eissa, Shimaa; Siaj, Mohamed; Zourob, Mohammed

    2015-07-15

    Brevetoxins (BTXs) are very potent marine neurotoxins that increased in geographical distribution in the past decade causing the illness clinically described as neurological shellfish poisoning (NSP). The ethical problems as well as the technical difficulties associated with the currently employed analysis methods for marine toxins are encouraging the research for suitable alternatives to be applied in a regulatory monitoring regime. Here, we report an electrochemical biosensor platform for BTX-2 detection utilising aptamer as specific receptor. Using in vitro selection, high affinity DNA aptamers to BTX-2 were successfully selected for the first time from a large pool of random sequences. The binding of BTX-2 to aptamer pools/clones was monitored using fluorescence and electrochemical impedance spectroscopy (EIS). The aptamer BT10 exhibited the highest binding affinity to BTX-2, with a dissociation constant of 42nM. The effects of the incubation time, pH and metal ions concentrations on the aptamer-toxin binding were studied. The aptamer BT10 was used to construct a label-free competitive impedimetric biosensor for BTX-2 achieving a detection limit of 106pg/ml. We observed a high degree of cross reactivity of the selected aptamer to the two similar congeners, BTX-2 and -3, whereas no cross reactivity to other marine toxins was obtained. Moreover, the aptasensor was applied for the detection of BTX-2 in spiked shellfish extract showing a very high recovery percentage. We believe that the proposed aptasensor will facilitate the routine detection of BTX-2 in food samples. PMID:25725463

  11. Electrochemical affinity biosensors for detection of mycotoxins: A review.

    PubMed

    Vidal, Juan C; Bonel, Laura; Ezquerra, Alba; Hernández, Susana; Bertolín, Juan R; Cubel, Carlota; Castillo, Juan R

    2013-11-15

    This review discusses the current state of electrochemical biosensors in the determination of mycotoxins in foods. Mycotoxins are highly toxic secondary metabolites produced by molds. The acute toxicity of these results in serious human and animal health problems, although it has been only since early 1960s when the first studied aflatoxins were found to be carcinogenic. Mycotoxins affect a broad range of agricultural products, most important cereals and cereal-based foods. A majority of countries, mentioning especially the European Union, have established preventive programs to control contamination and strict laws of the permitted levels in foods. Official methods of analysis of mycotoxins normally requires sophisticated instrumentation, e.g. liquid chromatography with fluorescence or mass detectors, combined with extraction procedures for sample preparation. For about sixteen years, the use of simpler and faster analytical procedures based on affinity biosensors has emerged in scientific literature as a very promising alternative, particularly electrochemical (i.e., amperometric, impedance, potentiometric or conductimetric) affinity biosensors due to their simplicity and sensitivity. Typically, electrochemical biosensors for mycotoxins use specific antibodies or aptamers as affinity ligands, although recombinant antibodies, artificial receptors and molecular imprinted polymers show potential utility. This article deals with recent advances in electrochemical affinity biosensors for mycotoxins and covers complete literature from the first reports about sixteen years ago. PMID:23743326

  12. Electrochemical Sensors and Biosensors Based on Nanomaterials and Nanostructures

    SciTech Connect

    Zhu, Chengzhou; Yang, Guohai; Li, He; Du, Dan; Lin, Yuehe

    2014-10-29

    We report that considerable attention has been devoted to the integration of recognition elements with electronic elements to develop electrochemical sensors and biosensors.Various electrochemical devices, such as amperometric sensors, electrochemical impedance sensors, and electrochemical luminescence sensors as well as photoelectrochemical sensors, provide wide applications in the detection of chemical and biological targets in terms of electrochemical change of electrode interfaces. Here, this review focuses on recent advances in electrochemical sensors and biosensors based on nanomaterials and nanostructures during 2013 to 2014. The aim of this effort is to provide the reader with a clear and concise view of new advances in areas ranging from electrode engineering, strategies for electrochemical signal amplification, and novel electroanalytical techniques used in the miniaturization and integration of the sensors. Moreover, the authors have attempted to highlight areas of the latest and significant development of enhanced electrochemical nanosensors and nanobiosensors that inspire broader interests across various disciplines. Electrochemical sensors for small molecules, enzyme-based biosensors, genosensors, immunosensors, and cytosensors are reviewed herein (Figure 1). Such novel advances are important for the development of electrochemical sensors that open up new avenues and methods for future research. In conclusion, we recommend readers interested in the general principles of electrochemical sensors and electrochemical methods to refer to other excellent literature for a broad scope in this area.(3, 4) However, due to the explosion of publications in this active field, we do not claim that this Review includes all of the published works in the past two years and we apologize to the authors of excellent work, which is unintentionally left out.

  13. Electrochemical Sensors and Biosensors Based on Nanomaterials and Nanostructures

    DOE PAGESBeta

    Zhu, Chengzhou; Yang, Guohai; Li, He; Du, Dan; Lin, Yuehe

    2014-10-29

    We report that considerable attention has been devoted to the integration of recognition elements with electronic elements to develop electrochemical sensors and biosensors.Various electrochemical devices, such as amperometric sensors, electrochemical impedance sensors, and electrochemical luminescence sensors as well as photoelectrochemical sensors, provide wide applications in the detection of chemical and biological targets in terms of electrochemical change of electrode interfaces. Here, this review focuses on recent advances in electrochemical sensors and biosensors based on nanomaterials and nanostructures during 2013 to 2014. The aim of this effort is to provide the reader with a clear and concise view of new advancesmore » in areas ranging from electrode engineering, strategies for electrochemical signal amplification, and novel electroanalytical techniques used in the miniaturization and integration of the sensors. Moreover, the authors have attempted to highlight areas of the latest and significant development of enhanced electrochemical nanosensors and nanobiosensors that inspire broader interests across various disciplines. Electrochemical sensors for small molecules, enzyme-based biosensors, genosensors, immunosensors, and cytosensors are reviewed herein (Figure 1). Such novel advances are important for the development of electrochemical sensors that open up new avenues and methods for future research. In conclusion, we recommend readers interested in the general principles of electrochemical sensors and electrochemical methods to refer to other excellent literature for a broad scope in this area.(3, 4) However, due to the explosion of publications in this active field, we do not claim that this Review includes all of the published works in the past two years and we apologize to the authors of excellent work, which is unintentionally left out.« less

  14. Enzyme catalytic amplification of miRNA-155 detection with graphene quantum dot-based electrochemical biosensor.

    PubMed

    Hu, Tianxing; Zhang, Le; Wen, Wei; Zhang, Xiuhua; Wang, Shengfu

    2016-03-15

    A specific and sensitive method was developed for quantitative detection of miRNA by integrating horseradish peroxidase (HRP)-assisted catalytic reaction with a simple electrochemical RNA biosensor. The electrochemical biosensor was constructed by a double-stranded DNA structure. The structure was formed by the hybridization of thiol-tethered oligodeoxynucleotide probes (capture DNA), assembled on the gold electrode surface, with target DNA and aminated indicator probe (NH2-DNA). After the construction of the double-stranded DNA structure, the activated carboxyl groups of graphene quantum dots (GQDs) assembled on NH2-DNA. GQDs were used as a new platform for HRP immobilization through noncovalent assembly. HRP modified biosensor can effectively catalyze the hydrogen peroxide (H2O2)-mediated oxidation of 3,3',5,5'-tetramethylbenzidine (TMB), accompanied by a change from colorless to blue in solution color and an increased electrochemical current signal. Due to GQDs and enzyme catalysis, the proposed biosensor could sensitively detect miRNA-155 from 1 fM to 100 pM with a detection limit of 0.14 fM. High performance of the biosensor is attributed to the large surface-to-volume ratio, excellent compatibility of GQDs. For these advantages, the proposed method holds great potential for analysis of other interesting tumor makers. PMID:26453906

  15. Electrochemical biosensor based on immobilized enzymes and redox polymers

    DOEpatents

    Skotheim, Terje A.; Okamoto, Yoshiyuki; Hale, Paul D.

    1992-01-01

    The present invention relates to an electrochemical enzyme biosensor for use in liquid mixtures of components for detecting the presence of, or measuring the amount of, one or more select components. The enzyme electrode of the present invention is comprised of an enzyme, an artificial redox compound covalently bound to a flexible polymer backbone and an electron collector.

  16. Direct application of gold nanoparticles to one-pot electrochemical biosensors.

    PubMed

    Chen, Guifang; Tong, Hui; Gao, Tao; Chen, Yangyang; Li, Genxi

    2014-11-01

    Gold nanoparticles (AuNPs) have been widely employed for the fabrication of electrochemical biosensors. In most cases, AuNPs are immobilized on the surface of an electrode, so they are difficult to be regenerated, making the use of the biosensor unfriendly. In this work, by adopting AuNPs directly as the electrolytes, we have developed a novel AuNPs-based electrochemical detection system. In brief, AuNPs-catalyzed oxidation of glucose is combined with a HRP-catalyzed reaction as well as an electrocatalytic reaction to compose cascade reactions in the electrolyte. Thus, the intensity of the electrocatalytic signals has quantitative relation with the concentration of glucose, and favors the sensitive detection of glucose. Furthermore, because the catalysis of AuNPs may be blocked under the interaction with single-stranded DNA and unblocked in the presence of a complementary sequence, detection of DNA and even single-nucleotide polymorphism can thereby been achieved. This one-pot detection system can be operated and regenerated very easily, since all the components are integrated in the electrolytes of AuNPs, and the unmodified electrode can be reused after being rinsed. This concept by integrating the advantages of sensitive electrochemical detection with the easy-to-operate nanocolloidal system may also promote the development of other kinds of electrochemical biosensors. PMID:25300210

  17. FIBER OPTIC BIOSENSOR FOR DNA DAMAGE

    EPA Science Inventory

    This paper describes a fiber optic biosensor for the rapid and sensitive detection of radiation-induced or chemically-induced oxidative DNA damage. The assay is based on the hybridization and temperature-induced dissociation (melting curves) of synthetic oligonucleotides. The...

  18. Bioelectrochemical interface engineering: toward the fabrication of electrochemical biosensors, biofuel cells, and self-powered logic biosensors.

    PubMed

    Zhou, Ming; Dong, Shaojun

    2011-11-15

    Over the past decade, researchers have devoted considerable attention to the integration of living organisms with electronic elements to yield bioelectronic devices. Not only is the integration of DNA, enzymes, or whole cells with electronics of scientific interest, but it has many versatile potential applications. Researchers are using these ideas to fabricate biosensors for analytical applications and to assemble biofuel cells (BFCs) and biomolecule-based devices. Other research efforts include the development of biocomputing systems for information processing. In this Account, we focus on our recent progress in engineering at the bioelectrochemical interface (BECI) for the rational design and construction of important bioelectronic devices, ranging from electrochemical (EC-) biosensors to BFCs, and self-powered logic biosensors. Hydrogels and sol-gels provide attractive materials for the immobilization of enzymes because they make EC-enzyme biosensors stable and even functional in extreme environments. We use a layer-by-layer (LBL) self-assembly technique to fabricate multicomponent thin films on the BECI at the nanometer scale. Additionally, we demonstrate how carbon nanomaterials have paved the way for new and improved EC-enzyme biosensors. In addition to the widely reported BECI-based electrochemical impedance spectroscopy (EIS)-type aptasensors, we integrate the LBL technique with our previously developed "solid-state probe" technique for redox probes immobilization on electrode surfaces to design and fabricate BECI-based differential pulse voltammetry (DPV)-type aptasensors. BFCs can directly harvest energy from ambient biofuels as green energy sources, which could lead to their application as simple, flexible, and portable power sources. Porous materials provide favorable microenvironments for enzyme immobilization, which can enhance BFC power output. Furthermore, by introducing aptamer-based logic systems to BFCs, such systems could be applied as self

  19. Development of an electrochemical biosensor for alkylphenol detection.

    PubMed

    Belkhamssa, Najet; da Costa, João P; Justino, Celine I L; Santos, Patrícia S M; Cardoso, Susana; Duarte, Armando C; Rocha-Santos, Teresa; Ksibi, Mohamed

    2016-09-01

    In this work, electrochemical biosensors based on field effect transistors (FET) with single-walled carbon nanotubes (SWCNT) were constructed as disposable analytical devices to detect alkylphenols through immunoreaction using 4-nonylphenol (NP) as model analyte, and validated by comparison with enzyme-linked immunosorbent assay (ELISA). The calibration curve displays a working range with five concentrations between 5 and 500µgL(-1), and for each concentration, five biosensors were analysed for reproducibility estimation and two analytical measurements were performed for each biosensor for repeatability estimation. The accuracy of the biosensors was validated by analyzing NP contents in ten spiked artificial seawater samples and comparing these results to those obtained with the traditional ELISA methodology. Excellent analytical performance was obtained with reproducibility of 0.56±0.08%, repeatability of 0.5±0.2%, limit of detection for NP as low as 5µgL(-1), and average recovery between 97.8% and 104.6%. This work demonstrates that simple biosensors can be used to detect hazardous priority substances in seawater samples, even at low concentrations. PMID:27343574

  20. Recent Development of Nano-Materials Used in DNA Biosensors

    PubMed Central

    Xu, Kai; Huang, Junran; Ye, Zunzhong; Ying, Yibin; Li, Yanbin

    2009-01-01

    As knowledge of the structure and function of nucleic acid molecules has increased, sequence-specific DNA detection has gained increased importance. DNA biosensors based on nucleic acid hybridization have been actively developed because of their specificity, speed, portability, and low cost. Recently, there has been considerable interest in using nano-materials for DNA biosensors. Because of their high surface-to-volume ratios and excellent biological compatibilities, nano-materials could be used to increase the amount of DNA immobilization; moreover, DNA bound to nano-materials can maintain its biological activity. Alternatively, signal amplification by labeling a targeted analyte with nano-materials has also been reported for DNA biosensors in many papers. This review summarizes the applications of various nano-materials for DNA biosensors during past five years. We found that nano-materials of small sizes were advantageous as substrates for DNA attachment or as labels for signal amplification; and use of two or more types of nano-materials in the biosensors could improve their overall quality and to overcome the deficiencies of the individual nano-components. Most current DNA biosensors require the use of polymerase chain reaction (PCR) in their protocols. However, further development of nano-materials with smaller size and/or with improved biological and chemical properties would substantially enhance the accuracy, selectivity and sensitivity of DNA biosensors. Thus, DNA biosensors without PCR amplification may become a reality in the foreseeable future. PMID:22346713

  1. Peptide biosensors for the electrochemical measurement of protein kinase activity.

    PubMed

    Kerman, Kagan; Song, Haifeng; Duncan, James S; Litchfield, David W; Kraatz, Heinz-Bernhard

    2008-12-15

    The kinase activities are elucidated using the novel redox-active cosubstrate adenosine 5'-[gamma-ferrocene] triphosphate (Fc-ATP), which enables the kinase-catalyzed transfer of a redox active gamma-phosphate-Fc to a hydroxyamino acid. In this report, a versatile electrochemical biosensor is developed for monitoring the activity and inhibition of a serine/threonine kinase, casein kinase 2 (CK2), and protein tyrosine kinases, Abl1-T315I and HER2, in buffered solutions and in cell lysates. The method is based on the labeling of a specific phosphorylation event with Fc, followed by electrochemical detection. The electrochemical response obtained from the "ferrocenylated" peptides enables monitoring the activity of the kinase and its substrate, as well as the inhibition of small molecule inhibitors on protein phosphorylation. Kinetic information was extracted from the electrochemical measurements for the determination of K(m) and V(m) values, which were in agreement with those previously reported. Kinase reactions were also performed in the presence of well-defined inhibitors of CK2, 4,5,6,7-tetrabromo-2-azabenzimidazole, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole, and E-3-(2,3,4,5-tetrabromophenyl)acrylic acid as well as the nonspecific kinase inhibitors, staurosporine and N-benzoylstaurosporine. On the basis of the dependency of the Fc signal on inhibitor concentration, K(i) of the inhibitors was estimated, which were also in agreement with the literature values. The performance of the biosensor was optimized including the kinase reaction, incubation with Fc-ATP, and the small molecule inhibitors. Peptide modified electrochemical biosensors are promising candidates for cost-effective in vitro kinase activity and inhibitor screening assays. PMID:18989981

  2. Detection of Interferon gamma using graphene and aptamer based FET-like electrochemical biosensor.

    PubMed

    Farid, Sidra; Meshik, Xenia; Choi, Min; Mukherjee, Souvik; Lan, Yi; Parikh, Devanshi; Poduri, Shripriya; Baterdene, Undarmaa; Huang, Ching-En; Wang, Yung Yu; Burke, Peter; Dutta, Mitra; Stroscio, Michael A

    2015-09-15

    One of the primary goals in the scientific community is the specific detection of proteins for the medical diagnostics and biomedical applications. Interferon-gamma (IFN-γ) is associated with the tuberculosis susceptibility, which is one of the major health problems globally. We have therefore developed a DNA aptamer-based electrochemical biosensor that is used for the detection of IFN-γ with high selectivity and sensitivity. A graphene monolayer-based FET-like structure is incorporated on a PDMS substrate with the IFN-γ aptamer attached to graphene. Addition of target molecule induces a change in the charge distribution in the electrolyte, resulting in increase in electron transfer efficiency that was actively sensed by monitoring the change in current from the device. Change in current appears to be highly sensitive to the IFN-γ concentrations ranging from nanomolar (nM) to micromolar (μM) range. The detection limit of our IFN-γ electrochemical biosensor is found to be 83 pM. Immobilization of aptamer on graphene surface is verified using unique structural approach by Atomic Force Microscopy. Such simple and sensitive electrochemical biosensor has potential applications in infectious disease monitoring, immunology and cancer research in the future. PMID:25919809

  3. A sensitive DNA biosensor fabricated from gold nanoparticles and graphene oxide on a glassy carbon electrode.

    PubMed

    Hajihosseini, Saeedeh; Nasirizadeh, Navid; Hejazi, Mohammad Saeid; Yaghmaei, Parichereh

    2016-04-01

    A sensitive electrochemical DNA biosensor was developed for Helicobacter pylori (H. pylori) detection using differential pulse voltammetry. Single-stranded DNA probe was immobilized on a graphene oxide/gold nanoparticles modified glassy carbon electrode (GO/AuNPs/GCE). A hybridization reaction was conducted with the target DNA and the immobilized DNA on the electrode surface. Oracet blue (OB) was selected for the first time as a redox indicator for amplifying the electrochemical signal of DNA. Enhanced sensitivity was achieved through combining the excellent electric conductivity of GO/AuNPs and the electroactivity of the OB. The DNA biosensor displayed excellent performance to demonstrate the differences between the voltammetric signals of the OB obtained from different hybridization samples (non-complementary, mismatch and complementary DNAs). The proposed biosensor has a linear range of 60.0-600.0 pM and a detection limit of 27.0 pM for detection of H. pylori. In addition, the biosensor have responded very well in the simulated real sample evaluations, signifying its potential to be used in future clinical detection of the H. pylori bacteria. PMID:26838878

  4. Novel immobilization techniques in the fabrication of efficient electrochemical biosensors

    NASA Astrophysics Data System (ADS)

    Alva, Shridhara; Marx, Kenneth A.; Samuelson, Lynne A.; Kumar, Jayant; Tripathy, Sukant K.; Kaplan, David L.

    1996-02-01

    The development of enzyme electrodes plays a major role in the performance of an electrochemical biosensor. In this paper, we describe two generic methods for efficient immobilization of enzymes or biomolecules at the electrode surface. These methods are based on physical entrapment of the enzymes during biochemical polymerization of phenols and electrochemical copolymerization of aromatic diamines with enzymes that are covalently coupled to the monomer. Both of these techniques have proven to be chemically mild and provide efficient polymer matrices for the fabrication of enzyme electrodes. Enzymes including horseradish peroxidase, alkaline phosphatase and glucose oxidase have been immobilized in these polymeric matrices and used for electrochemical as well as colorimetric detection of various substrates. Response times of the order of 5 - 10 seconds and sensitivities of the order of mM have been achieved with these electrodes. The use of these immobilization techniques towards the development of microelectrode arrays for multianalyte sensors is also discussed.

  5. Engineering the bioelectrochemical interface using functional nanomaterials and microchip technique toward sensitive and portable electrochemical biosensors.

    PubMed

    Jia, Xiaofang; Dong, Shaojun; Wang, Erkang

    2016-02-15

    Electrochemical biosensors have played active roles at the forefront of bioanalysis because they have the potential to achieve sensitive, specific and low-cost detection of biomolecules and many others. Engineering the electrochemical sensing interface with functional nanomaterials leads to novel electrochemical biosensors with improved performances in terms of sensitivity, selectivity, stability and simplicity. Functional nanomaterials possess good conductivity, catalytic activity, biocompatibility and high surface area. Coupled with bio-recognition elements, these features can amplify signal transduction and biorecognition events, resulting in highly sensitive biosensing. Additionally, microfluidic electrochemical biosensors have attracted considerable attention on account of their miniature, portable and low-cost systems as well as high fabrication throughput and ease of scaleup. For example, electrochemical enzymetic biosensors and aptamer biosensors (aptasensors) based on the integrated microchip can be used for portable point-of-care diagnostics and environmental monitoring. This review is a summary of our recent progress in the field of electrochemical biosensors, including aptasensors, cytosensors, enzymatic biosensors and self-powered biosensors based on biofuel cells. We presented the advantages that functional nanomaterials and microfluidic chip technology bring to the electrochemical biosensors, together with future prospects and possible challenges. PMID:26001888

  6. Biosensors based on DNA-Functionalized Graphene

    NASA Astrophysics Data System (ADS)

    Vishnubhotla, Ramya; Ping, Jinglei; Vrudhula, Amey; Johnson, A. T. Charlie

    Since its discovery, graphene has been used for sensing applications due to its outstanding electrical properties and biocompatibility. Here, we demonstrate the capabilities of field effect transistors (FETs) based on CVD-grown graphene functionalized with commercially obtained DNA oligomers and aptamers for detection of various biomolecular targets (e.g., complementary DNA and small molecule drug targets). Graphene FETs were created with a scalable photolithography process that produces arrays consisting of 50-100 FETs with a layout suitable for multiplexed detection of four molecular targets. FETs were characterized via AFM to confirm the presence of the aptamer. From the measured electrical characteristics, it was determined that binding of molecular targets by the DNA chemical recognition element led to a reproducible, concentration-dependent shift in the Dirac voltage. This biosensor class is potentially suitable for applications in drug detection. This work is funded by NIH through the Center for AIDS Research at the University of Pennsylvania.

  7. Design and characterization of novel electrochemical biosensors

    SciTech Connect

    Naser, N.

    1992-01-01

    This dissertation describes various novel approaches for fabricating fast responding, sensitive and selective biocatalytic sensors. One avenue is aimed at evaluating new plant tissues (rich with enzymatic activity in its own natural environment) within a carbon paste matrix or packed within the micropores of reticulated vitreous carbon. New self-supported open-tubular tissue bioreactors were also designed. The biocatalytic activity of these whole cells was employed for biosensing assays and for in-situ elimination of potential interferences via enzymatic consumption. Catalytic modified electrodes were developed to promote catalytic activity toward important analytes with a sluggish redox process. Such electrodes relied on the use of metal-dispersed carbon which lowered substantially the overvoltage. A new and rapid bulk modification scheme, based on incorporating the modifier within a rigid graphite-wax matrix was introduced. The new fabrication strategy was illustrated for the immobilization of electrocatalysts, enzymes and tissues. Effective organic-phase biosensors were developed and evaluated. They relied on the biocatalytic activity of Ta brockii alcohol dehydrogenase and of various plant tissues for rapid biosensing of secondary alcohols and phenolic compounds in non-aqueous media, respectively. Significant gains in the sensitivity of an alcohol sensor were obtained via biocatalytic preconcentration of intermediary products followed by a stripping measurement step. Peatmoss and Russian-thistle modified electrodes were developed for the preconcentration and quantitation of copper and gold ions from dilute solutions. These electrodes offered lower detection limits than algae-modified electrodes. Enzyme electrodes were fabricated by the physical entrapment of enzymes within a conducting polymer coating, onto band nanoelectrodes made of polyacrylonitrile films. The microdistribution of the enzyme in such coatings was investigated by scanning tunneling microscopy.

  8. Electrochemical enzymatic biosensors using carbon nanofiber nanoelectrode arrays

    NASA Astrophysics Data System (ADS)

    Li, Jun; Li, Yi-fen; Swisher, Luxi Z.; Syed, Lateef U.; Prior, Allan M.; Nguyen, Thu A.; Hua, Duy H.

    2012-10-01

    The reduction of electrode size down to nanometers could dramatically enhance detection sensitivity and temporal resolution. Nanoelectrode arrays (NEAs) are of particular interest for ultrasensitive biosensors. Here we report the study of two types of biosensors for measuring enzyme activities using NEAs fabricated with vertically aligned carbon nanofibers (VACNFs). VACNFs of ~100 nm in average diameter and 3-5 μm in length were grown on conductive substrates as uniform vertical arrays which were then encapsulated in SiO2 matrix leaving only the tips exposed. We demonstrate that such VACNF NEAs can be used in profiling enzyme activities through monitoring the change in electrochemical signals induced by enzymatic reactions to the peptides attached to the VACNF tip. The cleavage of the tetrapeptide with a ferrocene tag by a cancerrelated protease (legumain) was monitored with AC voltammetry. Real-time electrochemical impedance spectroscopy (REIS) was used for fast label-free detection of two reversible processes, i.e. phosphorylation by c-Src tyrosine kinase and dephosphorylation by protein tyrosine phosphatase 1B (PTP1B). The REIS data of phosphorylation were slow and unreliable, but those of dephosphorylation showed large and fast exponential decay due to much higher activity of phosphatase PTP1B. The kinetic data were analyzed with a heterogeneous Michaelis-Menten model to derive the "specificity constant" kcat/Km, which is 8.2x103 M-1s-1 for legumain and (2.1 ± 0.1) x 107 M-1s-1 for phosphatase (PTP1B), well consistent with literature. It is promising to develop VACNF NEA based electrochemical enzymatic biosensors as portable multiplex electronic techniques for rapid cancer diagnosis and treatment monitoring.

  9. A novel electrochemical detection method for aptamer biosensors.

    PubMed

    Bang, Gyeong Sook; Cho, Suhyeong; Kim, Byung-Gee

    2005-12-15

    A beacon aptamer-based biosensor for the detection of thrombin was developed using electrochemical transduction method. Gold surface was modified with a beacon aptamer covalently linked at 5'-terminus with a linker containing a primary aliphatic amine. Methylene blue (MB) was intercalated into the beacon sequence, and used as an electrochemical marker. When the beacon aptamer immobilized on gold surface encounters thrombin, the hairpin forming beacon aptamer is conformationally changed to release the intercalated MB, resulting a decrease in electrical current intensity in voltamogram. The peak signal of the MB is clearly decreased by the binding of thrombin onto the beacon aptamer. The linear range of the signal was observed between 0 and 50.8 nM of thrombin with 0.999 correlation factor. This method was able to linearly and selectively detect thrombin with a detection limit of 11 nM. PMID:16257654

  10. Electrochemical determination of levetiracetam by screen-printed based biosensors.

    PubMed

    Alonso-Lomillo, M Asunción; Domínguez-Renedo, Olga; Matos, Patricia; Arcos-Martínez, M Julia

    2009-02-01

    This work shows an easy and fast electrochemical method for Levetiracetam (LEV) determination, which is a novel antiepileptic. Most of the methods used up to now for its determination required a pre-treatment of the sample. It is shown here that the developed Peroxidase based biosensors avoid this kind of drawbacks. Screen-printed carbon electrodes have been used as transducers for the Peroxidase immobilization by pyrrole electropolymerization. Experimental variables that can affect LEV chronoamperometric response, such as hydrogen peroxide concentration, pH and applied potential, have been optimized in order to perform a selective LEV determination. Under these conditions, the performance of the biosensors has been tested. The residual standard deviation (RSD) of the slopes of different calibration curves was 9.77% (n=4 and alpha=0.05) for the reproducibility and 7.73% (n=4 and alpha=0.05) in the case of the repeatability. An average limit of detection of 9.81x10(-6) M (alpha=beta=0.05) was obtained. The biosensors have been finally applied to the determination of LEV in complex matrices, such as pharmaceutical drugs and spiked human plasma samples, yielding successful results. PMID:19059814

  11. Chemically modified flexible strips as electrochemical biosensors.

    PubMed

    Thota, Raju; Ganesh, V

    2014-09-21

    A flexible and disposable strip sensor for non-enzymatic glucose detection is demonstrated in this work. The strips are prepared by using chemical modification processes followed by a simple electroless deposition of copper. Essentially, polyester overhead projector (OHP) transparent films are modified with a monolayer of 3-aminopropyltrimethoxysilane (APTMS) and polyaniline (PANI) conducting polymer. Later, nanostructured copper is deposited onto this modified film. Scanning electron microscope (SEM) and X-ray diffraction (XRD) studies are used for the structural, morphological and crystallinity characterization of the modified films. Electrochemical techniques, namely cyclic voltammetry (CV) and chronoamperometry (CA), are employed for the non-enzymatic detection of glucose. These studies clearly reveal the formation of homogeneous, close-packed spherical Cu particles converged into uniform film that exhibits a good catalytic activity towards the oxidation of glucose. The Cu/PANI/APTMS/OHP sensor displays a remarkable enhancement in the oxidation current density, a very high sensitivity value of 2.8456 mA cm(-2) per mM, and a linear concentration range from 100 μM to 6.5 mM associated with glucose detection. Detection limit is estimated to be 5 μM and the response time of the sensor is determined to be less than 5 s. For comparison, similar studies are performed without PANI, namely Cu/APTMS/OHP films for glucose detection. In this case, a sensitivity value of 2.4457 mA cm(-2) per mM and a linear concentration range of 100 μM-3 mM are estimated. The higher performance characteristics observed in the case of Cu/PANI/APTMS/OHP are attributed to the synergistic effects of the conducting polymer acting as an electron facilitator and the nanostructured Cu films. These disposable, flexible and low-cost strip sensors have also been applied to the detection of glucose in clinical blood serum samples and the results obtained agree very well with the actual glucose

  12. Electrochemical biosensor for Ni(2+) detection based on a DNAzyme-CdSe nanocomposite.

    PubMed

    Yang, Ying; Yuan, Zheng; Liu, Xing-Pei; Liu, Qiao; Mao, Chang-Jie; Niu, He-Lin; Jin, Bao-Kang; Zhang, Sheng-Yi

    2016-03-15

    The detection and speciation analysis of metal-ion is very important for environmental monitoring. A novel electrochemical biosensor for Nickel(II) detection based on a DNAzyme-CdSe nanocomposite was developed. We firstly hybridized with capture probe (DNA1) and sequentially with DNA (DNA2) on the gold electrode. Then CdSe QDs were incorporated the specific recognition of DNA2 by covalent assembling. Upon addition of nickel ion into the above system, the substrate strand of the immobilized DNAzyme was catalytically cleaved by target Ni(2+), resulting in disassociation of the shorter DNA fragments containing CdSe QDs. The remaining CdSe QDs on the electrode surface detected by differential pulse anodic stripping voltammetry (DPASV). Under optimal conditions, the as-prepared sensor exhibited high sensitivity and fast response to Ni(2+) with the linear range from 20 nM to 0.2mM and a low detection limit of 6.67 nM. The prepared biosensor also shows good stability and good reproducibility and high selectivity toward target Ni(2+) against other metal ions because of highly specific Ni(2+)-dependent DNAzyme. Thus, our strategy has a good potential in the environment surveys. PMID:26385732

  13. Development of a multiarray biosensor for DNA diagnostics

    SciTech Connect

    Vo-Dinh, T.; Isola, N.; Alarie, J.P.; Landis, D.; Griffin, G.D.; Allison, S.

    1998-11-01

    This work involves the development and evaluation of a multiarray biosensor for DNA diagnostics. The evaluation of various system components developed for the biosensor is discussed. The DNA probes labeled with visible and near infrared (NIR) dyes are evaluated. The detection system uses a two-dimensional charge-coupled device (CCD). Examples of application of gene probes in DNA hybridization experiments and in biomedical diagnosis (detection of the p53 cancer gene) are presented to illustrate the usefulness and potential of the biosensor device.

  14. High specific surface gold electrode on polystyrene substrate: Characterization and application as DNA biosensor.

    PubMed

    Yang, Zhiliu; Liu, Yichen; Lu, Wei; Yuan, Qingpan; Wang, Wei; Pu, Qiaosheng; Yao, Bo

    2016-05-15

    In the past decades, many efforts have been made to improve the sensitivity and specificity of electrochemical DNA biosensors. However, it is still strongly required to develop disposable and reliable DNA biosensors for wide and practical application. In this article, we reported superior electrochemical properties of an integrated plastic-gold electrode (PGE) fabricated in-house by chemical plating on polystyrene substrate. PGEs were found having extremely high capacity of DNA immobilization compared with gold electrodes fabricated by standard sputtering based photolithography. Unique nano-structured surface was observed on PGEs through morphology techniques, which would to some extend give an explanation to higher capacity of DNA immobilization on PGEs. A probable mechanism of carboxylic acid produced on polystyrene substrate after exposure to UV irradiation was proposed and discussed for the first time. This biosensor was applied to detection and manipulate of DNA hybridization. Detection limit of 7.2×10(-11)M and 1-500nM of linearity range was obtained. PMID:26992524

  15. Biotin determination in food supplements by an electrochemical magneto biosensor.

    PubMed

    Kergaravat, Silvina V; Gómez, Gabriel A; Fabiano, Silvia N; Laube Chávez, Tamara I; Pividori, María I; Hernández, Silvia R

    2012-08-15

    An electrochemical magneto biosensor for the rapid determination of biotin in food samples is reported. The affinity reaction was performed on streptavidin-modified magnetic microbeads as a solid support in a direct competitive format. The biotinylated horseradish peroxidase enzyme (biotin-HRP) competes with free biotin in the sample for the binding sites of streptavidin on the magnetic microbeads. The modified magnetic beads were then easily captured by a magneto graphite-epoxy composite electrode and the electrochemical signal was based on the enzymatic activity of the HRP enzyme under the addition of H(2)O(2) as the substrate and o-phenilendiamine as cosubstrate. The response was electrochemically detected by square wave voltammetry. The limit of detection was 8.4×10(-8) mol L(--1) of biotin (20 μg L(--1)) with a dynamic range from 0.94 to 2.4×10(-7) mol L(--1). Biotin-fortified commercial dietary supplement and infant formula samples were evaluated obtaining good performances in the results. Total time of analysis was 40 min per 20 assays. PMID:22841112

  16. Preparation of Electrochemical Biosensor for Detection of Organophosphorus Pesticides

    PubMed Central

    Gothwal, Ashish; Beniwal, Puneet; Dhull, Vikas

    2014-01-01

    Polyvinyl chloride (PVC) can be used to develop reaction beaker which acts as electrochemical cell for the measurement of OP pesticides. Being chemically inert, corrosion resistant, and easy in molding to various shapes and size, PVC can be used for the immobilization of enzyme. Organophosphorus hydrolase was immobilized covalently onto the chemically activated inner surface of PVC beaker by using glutaraldehyde as a coupling agent. The carbon nanotubes paste working electrode was constructed for amperometric measurement at a potential of +0.8 V. The biosensor showed optimum response at pH 8.0 with incubation temperature of 40°C. Km and Imax for substrate (methyl parathion) were 322.58 µM and 1.1 µA, respectively. Evaluation study showed a correlation of 0.985, which was in agreement with the standard method. The OPH biosensor lost 50% of its initial activity after its regular use for 25 times over a period of 50 days when stored in 0.1 M sodium phosphate buffer, pH 8.0 at 4°C. No interference was observed by interfering species. PMID:25667593

  17. Electrochemical aptamer scaffold biosensors for detection of botulism and ricin toxins.

    PubMed

    Fetter, Lisa; Richards, Jonathan; Daniel, Jessica; Roon, Laura; Rowland, Teisha J; Bonham, Andrew J

    2015-10-21

    Protein toxins present considerable health risks, but detection often requires laborious analysis. Here, we developed electrochemical aptamer biosensors for ricin and botulinum neurotoxins, which display robust and specific signal at nanomolar concentrations and function in dilute serum. These biosensors may aid future efforts for the rapid diagnosis of toxins. PMID:26323568

  18. An Evolution Based Biosensor Receptor DNA Sequence Generation Algorithm

    PubMed Central

    Kim, Eungyeong; Lee, Malrey; Gatton, Thomas M.; Lee, Jaewan; Zang, Yupeng

    2010-01-01

    A biosensor is composed of a bioreceptor, an associated recognition molecule, and a signal transducer that can selectively detect target substances for analysis. DNA based biosensors utilize receptor molecules that allow hybridization with the target analyte. However, most DNA biosensor research uses oligonucleotides as the target analytes and does not address the potential problems of real samples. The identification of recognition molecules suitable for real target analyte samples is an important step towards further development of DNA biosensors. This study examines the characteristics of DNA used as bioreceptors and proposes a hybrid evolution-based DNA sequence generating algorithm, based on DNA computing, to identify suitable DNA bioreceptor recognition molecules for stable hybridization with real target substances. The Traveling Salesman Problem (TSP) approach is applied in the proposed algorithm to evaluate the safety and fitness of the generated DNA sequences. This approach improves efficiency and stability for enhanced and variable-length DNA sequence generation and allows extension to generation of variable-length DNA sequences with diverse receptor recognition requirements. PMID:22315543

  19. A microfluidic paper-based electrochemical biosensor array for multiplexed detection of metabolic biomarkers

    NASA Astrophysics Data System (ADS)

    Zhao, Chen; Thuo, Martin M.; Liu, Xinyu

    2013-10-01

    Paper-based microfluidic devices have emerged as simple yet powerful platforms for performing low-cost analytical tests. This paper reports a microfluidic paper-based electrochemical biosensor array for multiplexed detection of physiologically relevant metabolic biomarkers. Different from existing paper-based electrochemical devices, our device includes an array of eight electrochemical sensors and utilizes a handheld custom-made electrochemical reader (potentiostat) for signal readout. The biosensor array can detect several analytes in a sample solution and produce multiple measurements for each analyte from a single run. Using the device, we demonstrate simultaneous detection of glucose, lactate and uric acid in urine, with analytical performance comparable to that of the existing commercial and paper-based platforms. The paper-based biosensor array and its electrochemical reader will enable the acquisition of high-density, statistically meaningful diagnostic information at the point of care in a rapid and cost-efficient way.

  20. Papers Based Electrochemical Biosensors: From Test Strips to Paper-Based Microfluidics

    SciTech Connect

    Liu, Bingwen; Du, Dan; Hua, Xin; Yu, Xiao-Ying; Lin, Yuehe

    2014-05-08

    Papers based biosensors such as lateral flow test strips and paper-based microfluidic devices (or paperfluidics) are inexpensive, rapid, flexible, and easy-to-use analytical tools. An apparent trend in their detection is to interpret sensing results from qualitative assessment to quantitative determination. Electrochemical detection plays an important role in quantification. This review focuses on electrochemical (EC) detection enabled biosensors. The first part provides detailed examples in paper test strips. The second part gives an overview of paperfluidics engaging EC detections. The outlook and recommendation of future directions of EC enabled biosensors are discussed in the end.

  1. First paraben substituted cyclotetraphosphazene compounds and DNA interaction analysis with a new automated biosensor.

    PubMed

    Çiftçi, Gönül Yenilmez; Şenkuytu, Elif; İncir, Saadet Elif; Yuksel, Fatma; Ölçer, Zehra; Yıldırım, Tuba; Kılıç, Adem; Uludağ, Yıldız

    2016-06-15

    Cancer, as one of the leading causes of death in the world, is caused by malignant cell division and growth that depends on rapid DNA replication. To develop anti-cancer drugs this feature of cancer could be exploited by utilizing DNA-damaging molecules. To achieve this, the paraben substituted cyclotetraphosphazene compounds have been synthesized for the first time and their effect on DNA (genotoxicity) has been investigated. The conventional genotoxicity testing methods are laborious, take time and are expensive. Biosensor based assays provide an alternative to investigate this drug/compound DNA interactions. Here for the first time, a new, easy and rapid screening method has been used to investigate the DNA damage, which is based on an automated biosensor device that relies on the real-time electrochemical profiling (REP™) technology. Using both the biosensor based screening method and the in vitro biological assay, the compounds 9 and 11 (propyl and benzyl substituted cyclotetraphosphazene compounds, respectively), have resulted in higher DNA damage than the others with 65% and 80% activity reduction, respectively. PMID:26852202

  2. Graphene as a signal amplifier for preparation of ultrasensitive electrochemical biosensors

    PubMed Central

    Filip, Jaroslav; Kasák, Peter; Tkac, Jan

    2016-01-01

    Early diagnostics of diseases performed with minimal money and time consumption has become achievable due to recent advances in development of biosensors. These devices use biorecognition elements for selective interaction with an analyte and signal readout is obtained via different types of transducers. Operational characteristics of biosensors have been reported to improve substantially, when a diverse range of nanomaterials was employed. This review presents construction of electrochemical biosensors based on graphene, atomically thin 2D carbon crystals, which is currently intensively studied nanomaterial. The most attractive directions of graphene applications in biosensor preparation are discussed here including novel detection and amplification schemes exploiting graphene’s unique electrochemical, physical and chemical properties. The future of graphene-based biosensors is most likely bright, but there is still a lot of work to do to fulfill high expectations. PMID:27242391

  3. DNA Diagnostics: Nanotechnology-enhanced Electrochemical Detection of Nucleic Acids

    PubMed Central

    Wei, Fang; Lillehoj, Peter B.; Ho, Chih-Ming

    2010-01-01

    The detection of mismatched base pairs in DNA plays a crucial role in the diagnosis of genetic-related diseases and conditions, especially for early stage treatment. Among the various biosensors that have been employed for DNA detection, electrochemical sensors show great promise since they are capable of precise DNA recognition and efficient signal transduction. Advancements in micro- and nanotechnologies, specifically fabrication techniques and new nanomaterials, have enabled for the development of highly sensitive, highly specific sensors making them attractive for the detection of small sequence variations. Furthermore, the integration of sensors with sample preparation and fluidic processes enables for rapid, multiplexed DNA detection for point-of-care (POC) clinical diagnostics. PMID:20075759

  4. DNA-templated synthesis of PtAu bimetallic nanoparticle/graphene nanocomposites and their application in glucose biosensor

    NASA Astrophysics Data System (ADS)

    Leng, Jing; Wang, Wen-Min; Lu, Li-Min; Bai, Ling; Qiu, Xin-Lan

    2014-02-01

    In this paper, single-stranded DNA (ss-DNA) is demonstrated to functionalize graphene (GR) and to further guide the growth of PtAu bimetallic nanoparticles (PtAuNPs) on GR with high densities and dispersion. The obtained nanocomposites (PtAuNPs/ss-DNA/GR) were characterized by transmission electron microscopy (TEM), energy-dispersive X-ray spectrometer (EDS), and electrochemical techniques. Then, an enzyme nanoassembly was prepared by self-assembling glucose oxidase (GOD) on PtAuNP/ss-DNA/GR nanocomposites (GOD/PtAuNPs/ss-DNA/GR). The nanocomposites provided a suitable microenvironment for GOD to retain its biological activity. The direct and reversible electron transfer process between the active site of GOD and the modified electrode was realized without any extra electron mediator. Thus, the prepared GOD/PtAuNP/ss-DNA/GR electrode was proposed as a biosensor for the quantification of glucose. The effects of pH, applied potential, and temperature on the performance of the biosensor were discussed in detail and were optimized. Under optimal conditions, the biosensor showed a linearity with glucose concentration in the range of 1.0 to 1,800 μM with a detection limit of 0.3 μM (S/N = 3). The results demonstrate that the developed approach provides a promising strategy to improve the sensitivity and enzyme activity of electrochemical biosensors.

  5. DNA-templated synthesis of PtAu bimetallic nanoparticle/graphene nanocomposites and their application in glucose biosensor

    PubMed Central

    2014-01-01

    In this paper, single-stranded DNA (ss-DNA) is demonstrated to functionalize graphene (GR) and to further guide the growth of PtAu bimetallic nanoparticles (PtAuNPs) on GR with high densities and dispersion. The obtained nanocomposites (PtAuNPs/ss-DNA/GR) were characterized by transmission electron microscopy (TEM), energy-dispersive X-ray spectrometer (EDS), and electrochemical techniques. Then, an enzyme nanoassembly was prepared by self-assembling glucose oxidase (GOD) on PtAuNP/ss-DNA/GR nanocomposites (GOD/PtAuNPs/ss-DNA/GR). The nanocomposites provided a suitable microenvironment for GOD to retain its biological activity. The direct and reversible electron transfer process between the active site of GOD and the modified electrode was realized without any extra electron mediator. Thus, the prepared GOD/PtAuNP/ss-DNA/GR electrode was proposed as a biosensor for the quantification of glucose. The effects of pH, applied potential, and temperature on the performance of the biosensor were discussed in detail and were optimized. Under optimal conditions, the biosensor showed a linearity with glucose concentration in the range of 1.0 to 1,800 μM with a detection limit of 0.3 μM (S/N = 3). The results demonstrate that the developed approach provides a promising strategy to improve the sensitivity and enzyme activity of electrochemical biosensors. PMID:24572068

  6. Electrochemical biosensors based on nanofibres for cardiac biomarker detection: A comprehensive review.

    PubMed

    Rezaei, Babak; Ghani, Mozhdeh; Shoushtari, Ahmad Mousavi; Rabiee, Mohammad

    2016-04-15

    The vital importance of early and accurate diagnosis of cardiovascular diseases (CVDs) to prevent the irreversible damage or even death of patients has driven the development of biosensor devices for detection and quantification of cardiac biomarkers. Electrochemical biosensors offer rapid sensing, low cost, portability and ease of use. Over the past few years, nanotechnology has contributed to a tremendous improvement in the sensitivity of biosensors. In this review, the authors summarise the state-of-the-art of the application of one particular type of nanostructured material, i.e. nanofibres, for use in electrochemical biosensors for the ultrasensitive detection of cardiac biomarkers. A new way of classifying the nanofibre-based electrochemical biosensors according to the electrical conductance and the type of nanofibres is presented. Some key data from each article reviewed are highlighted, including the mechanism of detection, experimental conditions and the response range of the biosensor. The primary aim of this review is to emphasise the prospects for nanofibres for the future development of biosensors in diagnosis of CVDs as well as considering how to improve their characteristics for application in medicine. PMID:26657595

  7. Direct detection of DNA below ppb level based on thionin-functionalized layered MoS2 electrochemical sensors.

    PubMed

    Wang, Tanyuan; Zhu, Ruizhi; Zhuo, Junqiao; Zhu, Zhiwei; Shao, Yuanhua; Li, Meixian

    2014-12-16

    A layered MoS2-thionin composite was prepared by sonicating their mixture in an ionic liquid and gradient centrifugation. Because DNA is rarely present in single-stranded form, either naturally or after PCR amplification, the composite was used for fabrication of a double-stranded DNA (dsDNA) electrochemical biosensor due to stable electrochemical response, intercalation, and electrostatic interaction of thionin with DNA. The linear range over dsDNA concentration from 0.09 ng mL(-1) to 1.9 ng mL(-1) is obtained, and moreover, it is suitable for the detection of single-stranded DNA (ssDNA). The biosensor has been applied to the detection of circulating DNA from healthy human serum, and satisfactory results have been obtained. The constructed DNA electrochemical biosensor shows potential application in the fields of bioanalysis and clinic diagnosis. Furthermore, this work proposes a new method to construct electrochemical biosensors based on MoS2 sheets. PMID:25391335

  8. Electrochemical lectin based biosensors as a label-free tool in glycomics

    PubMed Central

    Bertók, Tomáš; Katrlík, Jaroslav; Gemeiner, Peter; Tkac, Jan

    2016-01-01

    Glycans and other saccharide moieties attached to proteins and lipids, or present on the surface of a cell, are actively involved in numerous physiological or pathological processes. Their structural flexibility (that is based on the formation of various kinds of linkages between saccharides) is making glycans superb “identity cards”. In fact, glycans can form more “words” or “codes” (i.e., unique sequences) from the same number of “letters” (building blocks) than DNA or proteins. Glycans are physicochemically similar and it is not a trivial task to identify their sequence, or - even more challenging - to link a given glycan to a particular physiological or pathological process. Lectins can recognise differences in glycan compositions even in their bound state and therefore are most useful tools in the task to decipher the “glycocode”. Thus, lectin-based biosensors working in a label-free mode can effectively complement the current weaponry of analytical tools in glycomics. This review gives an introduction into the area of glycomics and then focuses on the design, analytical performance, and practical utility of lectin-based electrochemical label-free biosensors for the detection of isolated glycoproteins or intact cells. PMID:27239071

  9. A liquid-crystal-based DNA biosensor for pathogen detection

    NASA Astrophysics Data System (ADS)

    Khan, Mashooq; Khan, Abdur Rahim; Shin, Jae-Ho; Park, Soo-Young

    2016-03-01

    A liquid-crystal (LC)-filled transmission electron microscopy (TEM) grid cell coated with the cationic surfactant dodecyltrimethylammonium bromide (DTAB), to which a single-stranded deoxyribonucleic acid probe (ssDNAprobe) was adsorbed at the LC/aqueous interface (TEMDTAB/DNA), was applied for the highly specific detection of target DNA molecules. The DTAB-coated E7 (used LC mixture) in the TEM grid (TEMDTAB) exhibited a homeotropic orientation, and changed to a planar orientation upon adsorption of the ssDNAprobe. The TEMDTAB/DNA was then exposed to complementary (target) ssDNA, which resulted in a planar-to-homeotropic configurational change of E7 that could be observed through a polarized optical microscope under crossed polarizers. The optimum adsorption density (2 μM) of ssDNAprobe enabled the detection of ≥0.05 nM complementary ssDNA. This TEMDTAB/DNA biosensor could differentiate complementary ssDNA from mismatched ssDNA as well as double-stranded DNA. It also successfully detected the genomic DNAs of the bacterium Erwinia carotovora and the fungi Rhazictonia solani. Owe to the high specificity, sensitivity, and label-free detection, this biosensor may broaden the applications of LC-based biosensors to pathogen detection.

  10. A liquid-crystal-based DNA biosensor for pathogen detection.

    PubMed

    Khan, Mashooq; Khan, Abdur Rahim; Shin, Jae-Ho; Park, Soo-Young

    2016-01-01

    A liquid-crystal (LC)-filled transmission electron microscopy (TEM) grid cell coated with the cationic surfactant dodecyltrimethylammonium bromide (DTAB), to which a single-stranded deoxyribonucleic acid probe (ssDNAprobe) was adsorbed at the LC/aqueous interface (TEMDTAB/DNA), was applied for the highly specific detection of target DNA molecules. The DTAB-coated E7 (used LC mixture) in the TEM grid (TEMDTAB) exhibited a homeotropic orientation, and changed to a planar orientation upon adsorption of the ssDNAprobe. The TEMDTAB/DNA was then exposed to complementary (target) ssDNA, which resulted in a planar-to-homeotropic configurational change of E7 that could be observed through a polarized optical microscope under crossed polarizers. The optimum adsorption density (2 μM) of ssDNAprobe enabled the detection of ≥0.05 nM complementary ssDNA. This TEMDTAB/DNA biosensor could differentiate complementary ssDNA from mismatched ssDNA as well as double-stranded DNA. It also successfully detected the genomic DNAs of the bacterium Erwinia carotovora and the fungi Rhazictonia solani. Owe to the high specificity, sensitivity, and label-free detection, this biosensor may broaden the applications of LC-based biosensors to pathogen detection. PMID:26940532

  11. A liquid-crystal-based DNA biosensor for pathogen detection

    PubMed Central

    Khan, Mashooq; Khan, Abdur Rahim; Shin, Jae-Ho; Park, Soo-Young

    2016-01-01

    A liquid-crystal (LC)-filled transmission electron microscopy (TEM) grid cell coated with the cationic surfactant dodecyltrimethylammonium bromide (DTAB), to which a single-stranded deoxyribonucleic acid probe (ssDNAprobe) was adsorbed at the LC/aqueous interface (TEMDTAB/DNA), was applied for the highly specific detection of target DNA molecules. The DTAB-coated E7 (used LC mixture) in the TEM grid (TEMDTAB) exhibited a homeotropic orientation, and changed to a planar orientation upon adsorption of the ssDNAprobe. The TEMDTAB/DNA was then exposed to complementary (target) ssDNA, which resulted in a planar-to-homeotropic configurational change of E7 that could be observed through a polarized optical microscope under crossed polarizers. The optimum adsorption density (2 μM) of ssDNAprobe enabled the detection of ≥0.05 nM complementary ssDNA. This TEMDTAB/DNA biosensor could differentiate complementary ssDNA from mismatched ssDNA as well as double-stranded DNA. It also successfully detected the genomic DNAs of the bacterium Erwinia carotovora and the fungi Rhazictonia solani. Owe to the high specificity, sensitivity, and label-free detection, this biosensor may broaden the applications of LC-based biosensors to pathogen detection. PMID:26940532

  12. Mimicking enzymatic effects of cytochrome P450 by an efficient biosensor for in vitro detection of DNA damage.

    PubMed

    Jalalvand, Ali R; Gholivand, Mohammad-Bagher; Goicoechea, Hector C; Skov, Thomas; Mansouri, Kamran

    2015-08-01

    A novel biosensor for detecting DNA damage induced by benzo(a)pyrene (BP) and its metabolite was presented in this work. The nafion-solubilized single wall carbon nanotubes-ionic liquid (SWCNTs-NA-IL) composite film was prepared and then horseradish peroxidase (HRP) and double-stranded DNA were alternately assembled on the composite film by the layer-by-layer method. The biosensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), differential pulse voltammetry (DPV), scanning electron microscopy (SEM) and computational methods. UV-vis spectrophotometry was also used to investigate DNA damage induced by BP and its metabolites in solution. The DNA biosensor was treated separately in BP, hydrogen peroxide (H2O2) and in their mixture, respectively. The EIS analysis showed a decrease in the charge transfer resistance at the DNA/HRP/SWCNTs-NA-IL/GCE incubated in a mixture of HRP and H2O2, because HRP in the presence of H2O2 could mimic enzymatic effects of cytochrome P450 (CYP450) to metabolize BP which could cause significant DNA damage and the exposed DNA bases reduced the electrostatic repulsion of the negatively charged redox probe and leads to Faradaic impedance changes. Finally, a novel biosensor for BP determination was developed and this method provided an indirect, and quantitative estimation of DNA damage in vitro. PMID:26038105

  13. Immobilization Techniques in the Fabrication of Nanomaterial-Based Electrochemical Biosensors: A Review

    PubMed Central

    Putzbach, William; Ronkainen, Niina J.

    2013-01-01

    The evolution of 1st to 3rd generation electrochemical biosensors reflects a simplification and enhancement of the transduction pathway. However, in recent years, modification of the transducer with nanomaterials has become increasingly studied and imparts many advantages. The sensitivity and overall performance of enzymatic biosensors has improved tremendously as a result of incorporating nanomaterials in their fabrication. Given the unique and favorable qualities of gold nanoparticles, graphene and carbon nanotubes as applied to electrochemical biosensors, a consolidated survey of the different methods of nanomaterial immobilization on transducer surfaces and enzyme immobilization on these species is beneficial and timely. This review encompasses modification of enzymatic biosensors with gold nanoparticles, carbon nanotubes, and graphene. PMID:23580051

  14. Designing new strategy for controlling DNA orientation in biosensors.

    PubMed

    Feng, Chao; Ding, Hong-ming; Ren, Chun-lai; Ma, Yu-qiang

    2015-01-01

    Orientation controllable DNA biosensors hold great application potentials in recognizing small molecules and detecting DNA hybridization. Though electric field is usually used to control the orientation of DNA molecules, it is also of great importance and significance to seek for other triggered methods to control the DNA orientation. Here, we design a new strategy for controlling DNA orientation in biosensors. The main idea is to copolymerize DNA molecules with responsive polymers that can show swelling/deswelling transitions due to the change of external stimuli, and then graft the copolymers onto an uncharged substrate. In order to highlight the responsive characteristic, we take thermo-responsive polymers as an example, and reveal multi-responsive behavior and the underlying molecular mechanism of the DNA orientation by combining dissipative particle dynamics simulation and molecular theory. Since swelling/deswelling transitions can be also realized by using other stimuli-responsive (like pH and light) polymers, the present strategy is universal, which can enrich the methods of controlling DNA orientation and may assist with the design of the next generation of biosensors. PMID:26400770

  15. Designing new strategy for controlling DNA orientation in biosensors

    PubMed Central

    Feng, Chao; Ding, Hong-ming; Ren, Chun-lai; Ma, Yu-qiang

    2015-01-01

    Orientation controllable DNA biosensors hold great application potentials in recognizing small molecules and detecting DNA hybridization. Though electric field is usually used to control the orientation of DNA molecules, it is also of great importance and significance to seek for other triggered methods to control the DNA orientation. Here, we design a new strategy for controlling DNA orientation in biosensors. The main idea is to copolymerize DNA molecules with responsive polymers that can show swelling/deswelling transitions due to the change of external stimuli, and then graft the copolymers onto an uncharged substrate. In order to highlight the responsive characteristic, we take thermo-responsive polymers as an example, and reveal multi-responsive behavior and the underlying molecular mechanism of the DNA orientation by combining dissipative particle dynamics simulation and molecular theory. Since swelling/deswelling transitions can be also realized by using other stimuli-responsive (like pH and light) polymers, the present strategy is universal, which can enrich the methods of controlling DNA orientation and may assist with the design of the next generation of biosensors. PMID:26400770

  16. Mediator-free electrochemical biosensor based on buckypaper with enhanced stability and sensitivity for glucose detection.

    PubMed

    Ahmadalinezhad, Asieh; Wu, Guosheng; Chen, Aicheng

    2011-12-15

    Here we report on a novel platform based on buckypaper for the design of high-performance electrochemical biosensors. Using glucose oxidase as a model enzyme, we constructed a biocompatible mediator-free biosensor and studied the potential effect of the buckypaper on the stability of the biosensor with both amperometry and FTIR spectroscopy. The results showed that the biosensor responses sensitively and selectively to glucose with a considerable functional lifetime of over 80 days. The fabricated enzymatic sensor detects glucose with a dynamic linear range of over 9 mM and a detection limit of 0.01 mM. To examine the efficiency of enzyme immobilization, the Michaelis-Menten constant (K(M)(app)) was calculated to be 4.67 mM. In addition, the fabricated electrochemical biosensor shows high selectivity; no amperometric response to the common interference species such as ascorbic acid, uric acid and acetamidophenol was observed. The facile and robust buckypaper-based platform proposed in this study opens the door for the design of high-performance electrochemical biosensors for medical diagnostics and environmental monitoring. PMID:22014621

  17. Electrochemical and optical biosensors based on nanomaterials and nanostructures: a review.

    PubMed

    Li, Ming; Li, Rui; Li, Chang Ming; Wu, Nianqiang

    2011-01-01

    Nanomaterials and nanostructures exhibit unique size-tunable and shape-dependent physicochemical properties that are different from those of bulk materials. Advances of nanomaterials and nanostructures open a new door to develop various novel biosensors. The present work has reviewed the recent progress in electrochemical, surface plasmon resonance (SPR), surface-enhanced Raman scattering (SERS) and fluorescent biosensors based on nanomaterials and nanostructures. An emphasis is put on the research that demonstrates how the performance of biosensors such as the limit of detection, sensitivity and selectivity is improved by the use of nanomaterials and nanostructures. PMID:21622273

  18. Application of DNA Hybridization Biosensor as a Screening Method for the Detection of Genetically Modified Food Components

    PubMed Central

    Tichoniuk, Mariusz; Ligaj, Marta; Filipiak, Marian

    2008-01-01

    An electrochemical biosensor for the detection of genetically modified food components is presented. The biosensor was based on 21-mer single-stranded oligonucleotide (ssDNA probe) specific to either 35S promoter or nos terminator, which are frequently present in transgenic DNA cassettes. ssDNA probe was covalently attached by 5′-phosphate end to amino group of cysteamine self-assembled monolayer (SAM) on gold electrode surface with the use of activating reagents – water soluble 1-ethyl-3(3′-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxy-sulfosuccinimide (NHS). The hybridization reaction on the electrode surface was detected via methylene blue (MB) presenting higher affinity to ssDNA probe than to DNA duplex. The electrode modification procedure was optimized using 19-mer oligoG and oligoC nucleotides. The biosensor enabled distinction between DNA samples isolated from soybean RoundupReady® (RR soybean) and non-genetically modified soybean. The frequent introduction of investigated DNA sequences in other genetically modified organisms (GMOs) give a broad perspectives for analytical application of the biosensor.

  19. DETECTION OF DNA DAMAGE USING A FIBEROPTIC BIOSENSOR

    EPA Science Inventory

    A rapid and sensitive fiber optic biosensor assay for radiation-induced DNA damage is reported. For this assay, a biotin-labeled capture oligonucleotide (38 mer) was immobilized to an avidin-coated quartz fiber. Hybridization of a dye-labeled complementary sequence was observed...

  20. Passive micromixers and organic electrochemical transistors for biosensor applications

    NASA Astrophysics Data System (ADS)

    Kanakamedala, Senaka Krishna

    Fluid handling at the microscale has greatly affected different fields such as biomedical, pharmaceutical, biochemical engineering and environmental monitoring due to its reduced reagent consumption, portability, high throughput, lower hardware cost and shorter analysis time compared to large devices. The challenges associated with mixing of fluids in microscale enabled us in designing, simulating, fabricating and characterizing various micromixers on silicon and flexible polyester substrates. The mixing efficiency was evaluated by injecting the fluids through the two inlets and collecting the sample at outlet. The images collected from the microscope were analyzed, and the absorbance of the color product at the outlet was measured to quantify the mixing efficacy. A mixing efficiency of 96% was achieved using a flexible disposable micromixer. The potential for low-cost processing and the device response tuning using chemical doping or synthesis opened doorways to use organic semiconductor devices as transducers in chemical and biological sensor applications. A simple, inexpensive organic electrochemical transistor (OECT) based on conducting polymer poly(3,4- ethyelenedioxythiphene) poly(styrene sulfonate) (PEDOT:PSS) was fabricated using a novel one step fabrication method. The developed transistor was used as a biosensor to detect glucose and glutamate. The developed glucose sensor showed a linear response for the glucose levels ranging from 1 muM-10 mM and showed a decent response for the glucose levels similar to those found in human saliva and to detect glutamate released from brain tumor cells. The developed glutamate sensor was used to detect the glutamate released from astrocytes and glioma cells after stimulation, and the results are compared with fluorescent spectrophotometer. The developed sensors employ simple fabrication, operate at low potentials, utilize lower enzyme concentrations, do not employ enzyme immobilization techniques, require only 5 muL of

  1. Microfabricated Electrochemical Cell-Based Biosensors for Analysis of Living Cells In Vitro

    PubMed Central

    Wang, Jun; Wu, Chengxiong; Hu, Ning; Zhou, Jie; Du, Liping; Wang, Ping

    2012-01-01

    Cellular biochemical parameters can be used to reveal the physiological and functional information of various cells. Due to demonstrated high accuracy and non-invasiveness, electrochemical detection methods have been used for cell-based investigation. When combined with improved biosensor design and advanced measurement systems, the on-line biochemical analysis of living cells in vitro has been applied for biological mechanism study, drug screening and even environmental monitoring. In recent decades, new types of miniaturized electrochemical biosensor are emerging with the development of microfabrication technology. This review aims to give an overview of the microfabricated electrochemical cell-based biosensors, such as microelectrode arrays (MEA), the electric cell-substrate impedance sensing (ECIS) technique, and the light addressable potentiometric sensor (LAPS). The details in their working principles, measurement systems, and applications in cell monitoring are covered. Driven by the need for high throughput and multi-parameter detection proposed by biomedicine, the development trends of electrochemical cell-based biosensors are also introduced, including newly developed integrated biosensors, and the application of nanotechnology and microfluidic technology. PMID:25585708

  2. Functionalized ensembles of nanoelectrodes as affinity biosensors for DNA hybridization detection.

    PubMed

    Silvestrini, Morena; Fruk, Ljiljana; Ugo, Paolo

    2013-02-15

    A novel electrochemical biosensor for DNA hybridization detection based on nanoelectrode ensembles (NEEs) is presented. NEEs are prepared by electroless deposition of gold into the pores of a templating track-etched polycarbonate (PC) membrane. The wide surface of the templating membrane surrounding the nanoelectrodes is exploited to bind the capture DNA probes via amide coupling with the carboxylic groups present on the PC surface. The probes are then hybridized with the complementary target labelled with glucose oxidase (GO(x)). The occurrence of the hybridization event is detected by adding, to the supporting electrolyte, excess glucose as the substrate and the (ferrocenylmethyl) trimethylammonium cation (FA(+)) as suitable redox mediator. In the case of positive hybridization, an electrocatalytic current is detected. In the proposed sensor, the biorecognition event and signal transduction occur in different but neighbouring sites, i.e., the PC surface and the nanoelectrodes, respectively; these sites are separated albeit in close proximity on a nanometer scale. Finally, the possibility to activate the PC surface by treatment with permanganate is demonstrated and the analytical performances of biosensors prepared with KMnO(4)-treated NEEs and native NEEs are compared and critically evaluated. The proposed biosensor displays high selectivity and sensitivity, with the capability to detect few picomoles of target DNA. PMID:22898659

  3. Electrochemical DNA Hybridization Sensors Based on Conducting Polymers

    PubMed Central

    Rahman, Md. Mahbubur; Li, Xiao-Bo; Lopa, Nasrin Siraj; Ahn, Sang Jung; Lee, Jae-Joon

    2015-01-01

    Conducting polymers (CPs) are a group of polymeric materials that have attracted considerable attention because of their unique electronic, chemical, and biochemical properties. This is reflected in their use in a wide range of potential applications, including light-emitting diodes, anti-static coating, electrochromic materials, solar cells, chemical sensors, biosensors, and drug-release systems. Electrochemical DNA sensors based on CPs can be used in numerous areas related to human health. This review summarizes the recent progress made in the development and use of CP-based electrochemical DNA hybridization sensors. We discuss the distinct properties of CPs with respect to their use in the immobilization of probe DNA on electrode surfaces, and we describe the immobilization techniques used for developing DNA hybridization sensors together with the various transduction methods employed. In the concluding part of this review, we present some of the challenges faced in the use of CP-based DNA hybridization sensors, as well as a future perspective. PMID:25664436

  4. Fabrication of Electrochemical Model Influenza A Virus Biosensor Based on the Measurements of Neuroaminidase Enzyme Activity.

    PubMed

    Anik, Ülkü; Tepeli, Yudum; Diouani, Mohamed F

    2016-06-21

    Neuroaminidase (NA) enzyme is a kind of glycoprotein that is found on the influenza A virus. During infection, NA is important for the release of influenza virions from the host cell surface together with viral aggregates. It may also be involved in targeting the virus to respiratory epithelial cells. In this study, a model electrochemical influenza A viral biosensor in which receptor-binding properties have been based on NA was developed for the first time. The biosensor's working principle is based on monitoring the interactions between fetuin A and NA enzyme. The assay was monitored step by step by using electrochemical impedance spectroscopy. PMID:27281347

  5. Detection of Aeromonas hydrophila DNA oligonucleotide sequence using a biosensor design based on Ceria nanoparticles decorated reduced graphene oxide and Fast Fourier transform square wave voltammetry.

    PubMed

    Jafari, Safiye; Faridbod, Farnoush; Norouzi, Parviz; Dezfuli, Amin Shiralizadeh; Ajloo, Davood; Mohammadipanah, Fatemeh; Ganjali, Mohammad Reza

    2015-10-01

    A new strategy was introduced for ssDNA immobilization on a modified glassy carbon electrode. The electrode surface was modified using polyaniline and chemically reduced graphene oxide decorated cerium oxide nanoparticles (CeO2NPs-RGO). A single-stranded DNA (ssDNA) probe was immobilized on the modified electrode surface. Fast Fourier transform square wave voltammetry (FFT-SWV) was applied as detection technique and [Ru(bpy)3](2+/3+) redox signal was used as electrochemical marker. The hybridization of ssDNA with its complementary target caused a dramatic decrease in [Ru(bpy)3](2+/3+) FFT-SW signal. The proposed electrochemical biosensor was able to detect Aeromonas hydrophila DNA oligonucleotide sequence encoding aerolysin protein. Under optimal conditions, the biosensor showed excellent selectivity toward complementary sequence in comparison with noncomplementary and two-base mismatch sequences. The dynamic linear range of this electrochemical DNA biosensor for detecting 20-mer oligonucleotide sequence of A. hydrophila was from 1 × 10(-15) to 1 × 10(-8) mol L(-1). The proposed biosensor was successfully applied for the detection of DNA extracted from A. hydrophila in fish pond water up to 0.01 μg mL(-1) with RSD of 5%. Besides, molecular docking was applied to consider the [Ru(bpy)3](2+/3+) interaction with ssDNA before and after hybridization. PMID:26454462

  6. Multicolor fluorescent biosensor for multiplexed detection of DNA.

    PubMed

    Hu, Rong; Liu, Tao; Zhang, Xiao-Bing; Huan, Shuang-Yan; Wu, Cuichen; Fu, Ting; Tan, Weihong

    2014-05-20

    Development of efficient methods for highly sensitive and rapid screening of specific oligonucleotide sequences is essential to the early diagnosis of serious diseases. In this work, an aggregated cationic perylene diimide (PDI) derivative was found to efficiently quench the fluorescence emission of a variety of anionic oligonucleotide-labeled fluorophores that emit at wavelengths from the visible to NIR region. This broad-spectrum quencher was then adopted to develop a multicolor biosensor via a label-free approach for multiplexed fluorescent detection of DNA. The aggregated perylene derivative exhibits a very high quenching efficiency on all ssDNA-labeled dyes associated with biosensor detection, having efficiency values of 98.3 ± 0.9%, 97 ± 1.1%, and 98.2 ± 0.6% for FAM, TAMRA, and Cy5, respectively. An exonuclease-assisted autocatalytic target recycling amplification was also integrated into the sensing system. High quenching efficiency combined with autocatalytic target recycling amplification afforded the biosensor with high sensitivity toward target DNA, resulting in a detection limit of 20 pM, which is about 50-fold lower than that of traditional unamplified homogeneous fluorescent assay methods. The quencher did not interfere with the catalytic activity of nuclease, and the biosensor could be manipulated in either preaddition or postaddition manner with similar sensitivity. Moreover, the proposed sensing system allows for simultaneous and multicolor analysis of several oligonucleotides in homogeneous solution, demonstrating its potential application in the rapid screening of multiple biotargets. PMID:24731194

  7. A ratiometric electrochemical biosensor for sensitive detection of Hg2+ based on thymine-Hg2+-thymine structure.

    PubMed

    Xiong, Erhu; Wu, Liang; Zhou, Jiawan; Yu, Peng; Zhang, Xiaohua; Chen, Jinhua

    2015-01-01

    In this paper, a simple, selective and reusable electrochemical biosensor for the sensitive detection of mercury ions (Hg(2+)) has been developed based on thymine (T)-rich stem-loop (hairpin) DNA probe and a dual-signaling electrochemical ratiometric strategy. The assay strategy includes both "signal-on" and "signal-off" elements. The thiolated methylene blue (MB)-modified T-rich hairpin DNA capture probe (MB-P) firstly self-assembled on the gold electrode surface via Au-S bond. In the presence of Hg(2+), the ferrocene (Fc)-labeled T-rich DNA probe (Fc-P) hybridized with MB-P via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. As a result, the hairpin MB-P was opened, the MB tags were away from the gold electrode surface and the Fc tags closed to the gold electrode surface. These conformation changes led to the decrease of the oxidation peak current of MB (IMB), accompanied with the increase of that of Fc (IFc). The logarithmic value of IFc/IMB is linear with the logarithm of Hg(2+) concentration in the range from 0.5 nM to 5000 nM, and the detection limit of 0.08 nM is much lower than 10nM (the US Environmental Protection Agency (EPA) limit of Hg(2+) in drinking water). What is more, the developed DNA-based electrochemical biosensor could be regenerated by adding cysteine and Mg(2+). This strategy provides a simple and rapid approach for the detection of Hg(2+), and has promising application in the detection of Hg(2+) in real environmental samples. PMID:25467465

  8. Enzyme-based electrochemical biosensors for determination of organophosphorus and carbamate pesticides

    SciTech Connect

    Everett, W.R.; Rechnitz, G.A.

    1999-01-01

    A mini review of enzyme-based electrochemical biosensors for inhibition analysis of organophosphorus and carbamate pesticides is presented. Discussion includes the most recent literature to present advances in detection limits, selectivity and real sample analysis. Recent reviews on the monitoring of pesticides and their residues suggest that the classical analytical techniques of gas and liquid chromatography are the most widely used methods of detection. These techniques, although very accurate in their determinations, can be quite time consuming and expensive and usually require extensive sample clean up and pro-concentration. For these and many other reasons, the classical techniques are very difficult to adapt for field use. Numerous researchers, in the past decade, have developed and made improvements on biosensors for use in pesticide analysis. This mini review will focus on recent advances made in enzyme-based electrochemical biosensors for the determinations of organophosphorus and carbamate pesticides.

  9. Recent Advances in Electrochemical Biosensors Based on Fullerene-C60 Nano-Structured Platforms

    PubMed Central

    Pilehvar, Sanaz; De Wael, Karolien

    2015-01-01

    Nanotechnology is becoming increasingly important in the field of (bio)sensors. The performance and sensitivity of biosensors is greatly improved with the integration of nanomaterials into their construction. Since its first discovery, fullerene-C60 has been the object of extensive research. Its unique and favorable characteristics of easy chemical modification, conductivity, and electrochemical properties has led to its tremendous use in (bio)sensor applications. This paper provides a concise review of advances in fullerene-C60 research and its use as a nanomaterial for the development of biosensors. We examine the research work reported in the literature on the synthesis, functionalization, approaches to nanostructuring electrodes with fullerene, and outline some of the exciting applications in the field of (bio)sensing. PMID:26610583

  10. A silicon nanowire-based electrochemical glucose biosensor with high electrocatalytic activity and sensitivity

    NASA Astrophysics Data System (ADS)

    Su, Shao; He, Yao; Song, Shiping; Li, Di; Wang, Lihua; Fan, Chunhai; Lee, Shuit-Tong

    2010-09-01

    An electrochemical glucose biosensor was developed by immobilizing glucose oxidase (GOx) on an electrode decorated with a novel nanostructure, silicon nanowires (SiNWs) with in situ grown gold nanoparticles (AuNPs). The immobilized GOx displayed a pair of well-defined and quasi-reversible redox peaks with a formal potential (E°') of -0.376 V in a phosphate buffer solution. The fabricated glucose biosensor has good electrocatalytic activity toward oxidation of glucose. In addition, such resultant SiNWs-based glucose biosensor possesses high biological affinity. Particularly, the apparent Michaelis-Mentan constant was estimated to be 0.902 mM, which is much smaller than the reported values for GOx at a range of nanomaterials-incorporated electrodes. Consequently, this novel SiNWs-based biosensor is expected to be a promising tool for biological assays (e.g., monitoring blood glucose).

  11. Conducting polymer based DNA biosensor for the detection of the Bacillus cereus group species

    NASA Astrophysics Data System (ADS)

    Velusamy, Vijayalakshmi; Arshak, Khalil; Korostynska, Olga; Oliwa, Kamila; Adley, Catherine

    2009-05-01

    Biosensor designs are emerging at a significant rate and play an increasingly important role in foodborne pathogen detection. Conducting polymers are excellent tools for the fabrication of biosensors and polypyrrole has been used in the detection of biomolecules due to its unique properties. The prime intention of this paper was to pioneer the design and fabrication of a single-strand (ss) DNA biosensor for the detection of the Bacillus cereus (B.cereus) group species. Growth of B. cereus, results in production of several highly active toxins. Therefore, consumption of food containing >106 bacteria/gm may results in emetic and diarrhoeal syndromes. The most common source of this bacterium is found in liquid food products, milk powder, mixed food products and is of particular concern in the baby formula industry. The electrochemical deposition technique, such as cyclic voltammetry, was used to develop and test a model DNA-based biosensor on a gold electrode electropolymerized with polypyrrole. The electrically conducting polymer, polypyrrole is used as a platform for immobilizing DNA (1μg) on the gold electrode surface, since it can be more easily deposited from neutral pH aqueous solutions of pyrrolemonomers. The average current peak during the electrodeposition event is 288μA. There is a clear change in the current after hybridization of the complementary oligonucleotide (6.35μA) and for the noncomplementary oligonucleotide (5.77μA). The drop in current after each event was clearly noticeable and it proved to be effective.

  12. Electronically type-sorted carbon nanotube-based electrochemical biosensors with glucose oxidase and dehydrogenase.

    PubMed

    Muguruma, Hitoshi; Hoshino, Tatsuya; Nowaki, Kohei

    2015-01-14

    An electrochemical enzyme biosensor with electronically type-sorted (metallic and semiconducting) single-walled carbon nanotubes (SWNTs) for use in aqueous media is presented. This research investigates how the electronic types of SWNTs influence the amperometric response of enzyme biosensors. To conduct a clear evaluation, a simple layer-by-layer process based on a plasma-polymerized nano thin film (PPF) was adopted because a PPF is an inactive matrix that can form a well-defined nanostructure composed of SWNTs and enzyme. For a biosensor with the glucose oxidase (GOx) enzyme in the presence of oxygen, the response of a metallic SWNT-GOx electrode was 2 times larger than that of a semiconducting SWNT-GOx electrode. In contrast, in the absence of oxygen, the response of the semiconducting SWNT-GOx electrode was retained, whereas that of the metallic SWNT-GOx electrode was significantly reduced. This indicates that direct electron transfer occurred with the semiconducting SWNT-GOx electrode, whereas the metallic SWNT-GOx electrode was dominated by a hydrogen peroxide pathway caused by an enzymatic reaction. For a biosensor with the glucose dehydrogenase (GDH; oxygen-independent catalysis) enzyme, the response of the semiconducting SWNT-GDH electrode was 4 times larger than that of the metallic SWNT-GDH electrode. Electrochemical impedance spectroscopy was used to show that the semiconducting SWNT network has less resistance for electron transfer than the metallic SWNT network. Therefore, it was concluded that semiconducting SWNTs are more suitable than metallic SWNTs for electrochemical enzyme biosensors in terms of direct electron transfer as a detection mechanism. This study makes a valuable contribution toward the development of electrochemical biosensors that employ sorted SWNTs and various enzymes. PMID:25522366

  13. Carbon nanotubes (CNTs) for the development of electrochemical biosensors.

    PubMed

    Lin, Yuehe; Yantasee, Wassana; Wang, Joseph

    2005-01-01

    Carbon nanotube (CNT) is a very attractive material for the development of biosensors because of its capability to provide strong electrocatalytic activity and minimize surface fouling of the sensors. This article reviews our recent developments of oxidase- and dehydrogenase-amperometric biosensors based on the immobilization of CNTs, the co-immobilization of enzymes on the CNTs/Nafion or the CNT/Teflon composite materials, or the attachment of enzymes on the controlled-density aligned CNT-nanoelectrode arrays. The excellent electrocatalytic activities of the CNTs on the redox reactions of hydrogen peroxide, nicotinamide adenine dinucleotide (NADH), and homocysteine have been demonstrated. Successful applications of the CNT-based biosensors reviewed herein include the low-potential detections of glucose, organophosphorus compounds, and alcohol. PMID:15574386

  14. Carbon Nanotubes (CNTs) for the Development of Electrochemical Biosensors

    SciTech Connect

    Lin, Yuehe; Yantasee, Wassana; Wang, Joseph

    2005-01-01

    Carbon nanotube (CNT) is a very attractive material for the development of biosensors because of its capability to provide strong electrocatalytic activity and minimize surface fouling of the sensors. This article reviews our recent developments of oxidase- and dehydrogenase-amperometric biosensors based on the immobilization of CNTs, the co-immobilization of enzymes on the CNTs/Nafion or the CNT/Teflon composite materials, or the attachment of enzymes on the controlled-density aligned CNT-nanoelectrode arrays. The excellent electrocatalytic activities of the CNTs on the redox reactions of hydrogen peroxide, nicotinamide adenine dinucleotide (NADH), and homocysteine have been demonstrated. Successful applications of the CNT-based biosensors reviewed herein include the low-potential detections of glucose, organophosphorus compounds, and alcohol.

  15. Ionic liquid of a gold nanocluster: a versatile matrix for electrochemical biosensors.

    PubMed

    Kwak, Kyuju; Kumar, S Senthil; Pyo, Kyunglim; Lee, Dongil

    2014-01-28

    Ionic liquids are room-temperature molten salts that are increasingly used in electrochemical devices, such as batteries, fuel cells, and sensors, where their intrinsic ionic conductivity is exploited. Here we demonstrate that combining anionic, redox-active Au25 clusters with imidazolium cations leads to a stable ionic liquid possessing both ionic and electronic conductivity. The Au25 ionic liquid was found to act as a versatile matrix for amperometric enzyme biosensors toward the detection of glucose. Enzyme electrodes prepared by incorporating glucose oxidase in the Au25 ionic liquid show high electrocatalytic activity and substrate affinity. Au25 clusters in the electrode were found to act as effective redox mediators as well as electronic conductors determining the detection sensitivity. With the unique electrochemical properties and almost unlimited structural tunability, the ionic liquids of quantum-sized gold clusters may serve as versatile matrices for a variety of electrochemical biosensors. PMID:24350837

  16. Characterization of immobilization methods of antiviral antibodies in serum for electrochemical biosensors

    NASA Astrophysics Data System (ADS)

    Huy, Tran Quang; Hanh, Nguyen Thi Hong; Van Chung, Pham; Anh, Dang Duc; Nga, Phan Thi; Tuan, Mai Anh

    2011-06-01

    In this paper, we describes different methods to immobilize Japanese encephalitis virus (JEV) antibodies in human serum onto the interdigitated surface of a microelectrode sensor for optimizing electrochemical detection: (1) direct covalent binding to the silanized surface, (2) binding to the silanized surface via a cross-linker of glutaraldehyde (GA), (3) binding to glutaraldehyde/silanized surface via goat anti-human IgG polyclonal antibody and (4) binding to glutaraldehyde/silanized surface via protein A (PrA). Field emission scanning electron microscopy, Fourier transform infrared spectrometry, and fluorescence microscopy are used to verify the characteristics of antibodies on the interdigitated surface after the serum antibodies immobilization. The analyzed results indicate that the use of protein A is an effective choice for immobilization and orientation of antibodies in serum for electrochemical biosensors. This study provides an advantageous immobilization method of serum containing antiviral antibodies to develop electrochemical biosensors for preliminary screening of viruses in clinical samples from outbreaks.

  17. A Review on the Electrochemical Sensors and Biosensors Composed of Nanowires as Sensing Material

    PubMed Central

    Yogeswaran, Umasankar; Chen, Shen-Ming

    2008-01-01

    The development and application of nanowires for electrochemical sensors and biosensors are reviewed in this article. Next generation sensor platforms will require significant improvements in sensitivity, specificity and parallelism in order to meet the future needs in variety of fields. Sensors made of nanowires exploit some fundamental nanoscopic effect in order to meet these requirements. Nanowires are new materials, which have the characteristic of low weight with extraordinary mechanical, electrical, thermal and multifunctional properties. The advantages such as size scale, aspect ratio and other properties of nanowires are especially apparent in the use of electrical sensors such as electrochemical sensors and in the use of field-effect transistors. The preparation methods of nanowires and their properties are discussed along with their advantages towards electrochemical sensors and biosensors. Some key results from each article are summarized, relating the concept and mechanism behind each sensor, with experimental conditions as well as their behavior at different conditions.

  18. A novel GMO biosensor for rapid ultrasensitive and simultaneous detection of multiple DNA components in GMO products.

    PubMed

    Huang, Lin; Zheng, Lei; Chen, Yinji; Xue, Feng; Cheng, Lin; Adeloju, Samuel B; Chen, Wei

    2015-04-15

    Since the introduction of genetically modified organisms (GMOs), there has been on-going and continuous concern and debates on the commercialization of products derived from GMOs. There is an urgent need for development of highly efficient analytical methods for rapid and high throughput screening of GMOs components, as required for appropriate labeling of GMO-derived foods, as well as for on-site inspection and import/export quarantine. In this study, we describe, for the first time, a multi-labeling based electrochemical biosensor for simultaneous detection of multiple DNA components of GMO products on the same sensing interface. Two-round signal amplification was applied by using both an exonuclease enzyme catalytic reaction and gold nanoparticle-based bio-barcode related strategies, respectively. Simultaneous multiple detections of different DNA components of GMOs were successfully achieved with satisfied sensitivity using this electrochemical biosensor. Furthermore, the robustness and effectiveness of the proposed approach was successfully demonstrated by application to various GMO products, including locally obtained and confirmed commercial GMO seeds and transgenetic plants. The proposed electrochemical biosensor demonstrated unique merits that promise to gain more interest in its use for rapid and on-site simultaneous multiple screening of different components of GMO products. PMID:25497983

  19. Gold nanoparticle based signal enhancement liquid crystal biosensors for DNA hybridization assays.

    PubMed

    Yang, Shengyuan; Liu, Yanmei; Tan, Hui; Wu, Chao; Wu, Zhaoyang; Shen, Guoli; Yu, Ruqin

    2012-03-18

    A novel signal enhanced liquid crystal biosensor based on using AuNPs for highly sensitive DNA detection has been developed. This biosensor not only significantly decreases the detection limit, but also offers a simple detection process and shows a good selectivity to distinguish perfectly matched target DNA from two-base mismatched DNA. PMID:22302154

  20. A Graphene and Aptamer Based Liquid Gated FET-Like Electrochemical Biosensor to Detect Adenosine Triphosphate.

    PubMed

    Mukherjee, Souvik; Meshik, Xenia; Choi, Min; Farid, Sidra; Datta, Debopam; Lan, Yi; Poduri, Shripriya; Sarkar, Ketaki; Baterdene, Undarmaa; Huang, Ching-En; Wang, Yung Yu; Burke, Peter; Dutta, Mitra; Stroscio, Michael A

    2015-12-01

    Here we report successful demonstration of a FET-like electrochemical nano-biosensor to accurately detect ultralow concentrations of adenosine triphosphate. As a 2D material, graphene is a promising candidate due to its large surface area, biocompatibility, and demonstrated surface binding chemistries and has been employed as the conducting channel. A short 20-base DNA aptamer is used as the sensing element to ensure that the interaction between the analyte and the aptamer occurs within the Debye length of the electrolyte (PBS). Significant increase in the drain current with progressive addition of ATP is observed whereas for control experiments, no distinct change in the drain current occurs. The sensor is found to be highly sensitive in the nanomolar (nM) to micromolar ( μM) range with a high sensitivity of 2.55 μA (mM) (-1), a detection limit as low as 10 pM, and it has potential application in medical and biological settings to detect low traces of ATP. This simplistic design strategy can be further extended to efficiently detect a broad range of other target analytes. PMID:26595926

  1. New Catalytic DNA Biosensors for Radionuclides and Metal ion

    SciTech Connect

    Yi Lu

    2008-03-01

    We aim to develop new DNA biosensors for simultaneous detection and quantification of bioavailable radionuclides, such as uranium, technetium, and plutonium, and metal contaminants, such as lead, chromium, and mercury. The sensors will be highly sensitive and selective. They will be applied to on-site, real-time assessment of concentration, speciation, and stability of the individual contaminants before and during bioremediation, and for long-term monitoring of DOE contaminated sites. To achieve this goal, we have employed a combinatorial method called “in vitro selection” to search from a large DNA library (~ 1015 different molecules) for catalytic DNA molecules that are highly specific for radionuclides or other metal ions through intricate 3-dimensional interactions as in metalloproteins. Comprehensive biochemical and biophysical studies have been performed on the selected DNA molecules. The findings from these studies have helped to elucidate fundamental principles for designing effective sensors for radionuclides and metal ions. Based on the study, the DNA have been converted to fluorescent or colorimetric sensors by attaching to it fluorescent donor/acceptor pairs or gold nanoparticles, with 11 part-per-trillion detection limit (for uranium) and over million fold selectivity (over other radionuclides and metal ions tested). Practical application of the biosensors for samples from the Environmental Remediation Sciences Program (ERSP) Field Research Center (FRC) at Oak Ridge has also been demonstrated.

  2. DNA microdevice for electrochemical detection of Escherichia coli 0157:H7 molecular markers.

    PubMed

    Berganza, J; Olabarria, G; García, R; Verdoy, D; Rebollo, A; Arana, S

    2007-04-15

    An electrochemical DNA sensor based on the hybridization recognition of a single-stranded DNA (ssDNA) probe immobilized onto a gold electrode to its complementary ssDNA is presented. The DNA probe is bound on gold surface electrode by using self-assembled monolayer (SAM) technology. An optimized mixed SAM with a blocking molecule preventing the nonspecific adsorption on the electrode surface has been prepared. In this paper, a DNA biosensor is designed by means of the immobilization of a single stranded DNA probe on an electrochemical transducer surface to recognize specifically Escherichia coli (E. coli) 0157:H7 complementary target DNA sequence via cyclic voltammetry experiments. The 21 mer DNA probe including a C6 alkanethiol group at the 5' phosphate end has been synthesized to form the SAM onto the gold surface through the gold sulfur bond. The goal of this paper has been to design, characterise and optimise an electrochemical DNA sensor. In order to investigate the oligonucleotide probe immobilization and the hybridization detection, experiments with different concentration of DNA and mismatch sequences have been performed. This microdevice has demonstrated the suitability of oligonucleotide Self-assembled monolayers (SAMs) on gold as immobilization method. The DNA probes deposited on gold surface have been functional and able to detect changes in bases sequence in a 21-mer oligonucleotide. PMID:17084071

  3. An Electrochemical DNA Microbiosensor Based on Succinimide-Modified Acrylic Microspheres

    PubMed Central

    Ulianas, Alizar; Heng, Lee Yook; Hanifah, Sharina Abu; Ling, Tan Ling

    2012-01-01

    An electrochemical microbiosensor for DNA has been fabricated based on new acrylic microspheres modified with reactive N-acryloxysuccinimide (NAS) functional groups. Hydrophobic poly(n-butylacrylate-N-acryloxysuccinimide) microspheres were synthesized in an emulsion form with a simple one-step photopolymerization technique. Aminated DNA probe was attached to the succinimde functional group of the acrylic microspheres via covalent bonding. The hybridization of the immobilized DNA probe with the complementary DNA was studied by differential pulse voltametry using anthraquninone-2-sulfonic acid monohydrate sodium salt (AQMS) as the electroactive hybridization label. The influences of many factors such as duration of DNA probe immobilization and hybridization, pH, type of ions, buffer concentrations, ionic strength, operational temperature and non-complementary DNA on the biosensor performance were evaluated. Under optimized conditions, the DNA microbiosensor demonstrated a linear response range to target DNA over a wide concentration range of 1.0 × 10−16 and 1.0 × 10−8 M with a lower limit of detection (LOD) of 9.46 × 10−17 M (R2 = 0.97). This DNA microbiosensor showed good reproducibility with 2.84% RSD (relative standard deviation) (n = 3). Application of the NAS-modified acrylic microspheres in the construction of DNA microbiosensor had improved the overall analytical performance of the resultant DNA microbiosensor when compared with other reported DNA biosensors using other nano-materials for membranes and microspheres as DNA immobilization matrices. PMID:22778594

  4. Electrochemical biosensor based on CdS nanostructure surfaces.

    PubMed

    Qian, Jiqing; Yan, Shancheng; Xiao, Zhongdang

    2012-01-15

    Well-defined hexangularly faced CdS nanorod arrays have been grown directly on a conductive ITO glass via a facile one-step and non-template hydrothermal approach. Gold nanoparticles were decorated onto the nanorods to enhance the electron transfer process of electrode. Glucose oxidase (GOD) was then immobilized on the CdS through crosslinking with chitosan (CS), which resulted in a glucose biosensor with high enzyme loading and excellent sensitivity. Such a chitosan-encapsulated GOD-based biosensor revealed a relatively rapid response time of less than 50s, and an approximate linear detection range of glucose concentration, from 50 to 500 μmol L(-1) with a detection limit of 38 μmol L(-1) and an electrode sensitivity of 5.9 μA mM(-1). PMID:22018624

  5. Increased electrocatalyzed performance through hairpin oligonucleotide aptamer-functionalized gold nanorods labels and graphene-streptavidin nanomatrix: Highly selective and sensitive electrochemical biosensor of carcinoembryonic antigen.

    PubMed

    Wen, Wei; Huang, Jing-Yi; Bao, Ting; Zhou, Jun; Xia, Hong-Xing; Zhang, Xiu-Hua; Wang, Sheng-Fu; Zhao, Yuan-Di

    2016-09-15

    We report a triplex signal amplification strategy for sensitive biosensing of cancer biomarker by taking advantage of hairpin-shaped oligonucleotide-functionalized gold nanorods (HO-GNRs), graphene and the avidin-biotin reation. The strategy expands electrochemical detection of carcinoembryonic antigen (CEA) by using an aptamer as biosensor's recognition element and HO-GNRs as signal enhancer. To construct this biosensor, the GNR was used as a carrier of horseradish peroxidase (HRP) and HO aptamer with a biotin at the 3'-end and a thiol at the 5'-end, which amplified the electrochemical response because of a large molar ratio of HRP to HO. In the presence of target CEA, the binding reactions of CEA with the loop portions of the HOs caused HOs' loop-stem structure opened and exposed the biotins, and then HRP-GNRs-HO conjugates were captured on graphene and streptavidin modified electrodes via the reaction between the exposed biotins and preimmobilized streptavidins. The accumulation of HRP effectively catalyzed the hydrogen peroxide-mediated oxidation of o-phenylenediamine to generate an electrochemical reduction current for CEA detection. Under optimal conditions, the electrochemical biosensor exhibited a wide dynamic range of 5pgmL(-1) and 50ngmL(-1) toward CEA standards with a low detection limit of 1.5pgmL(-1) (signal-to-noise ratio of 3). The proposed biosensor accurately detected CEA concentration in 8 human serum samples from patients with lung diseases, showing excellent correlations with standard chemiluminescence immunoassay. Furthermore, these results of target DNA detection made it abundantly clear that the proposed strategy can also be extended for detection of other relative biomarkers using different functional DNA structures, which shows great prospects in single-nucleotide polymorphisms analysis, biomedical sensing and application for accurate clinical diseases diagnostic. PMID:27111123

  6. Rapid and simple preparation of a reagentless glucose electrochemical biosensor.

    PubMed

    Zheng, Dan; Vashist, Sandeep Kumar; Al-Rubeaan, Khalid; Luong, John H T; Sheu, Fwu-Shan

    2012-08-21

    A rapid and simple procedure was developed for the preparation of a highly stable and leach-proof glucose oxidase (GOx)-bound glassy carbon electrode (GCE). Crosslinked GOx via glutaraldehyde was drop-cast on a KOH-pretreated GCE followed by drop-casting of 3-aminopropyltriethoxysilane (APTES) to form a stable bioactive layer. At -0.45 V, the biosensor exhibited a wide dynamic detection range of 0.5-48 mM for commercial glucose and 1.3-28.2 mM for Sugar-Chex blood glucose linearity standards. Several endogenous electroactive substances and drug metabolites commonly found in blood were tested and provoked no signal response. To our knowledge, the developed procedure is the most rapid method for preparing a glucose biosensor. The biosensor suffered no biofouling after 7 days of immersion in Sugar-Chex blood glucose. With excellent production reproducibility, GOx-bound electrodes stored dry at room temperature retained their initial activity after several weeks. PMID:22763782

  7. Acetylcholinesterase Biosensors for Electrochemical Detection of Organophosphorus Compounds: A Review

    PubMed Central

    Dhull, Vikas; Gahlaut, Anjum; Dilbaghi, Neeraj

    2013-01-01

    The exponentially growing population, with limited resources, has exerted an intense pressure on the agriculture sector. In order to achieve high productivity the use of pesticide has increased up to many folds. These pesticides contain organophosphorus (OP) toxic compounds which interfere with the proper functioning of enzyme acetylcholinesterase (AChE) and finally affect the central nervous system (CNS). So, there is a need for routine, continuous, on spot detection of OP compounds which are the main limitations associated with conventional analytical methods. AChE based enzymatic biosensors have been reported by researchers as the most promising tool for analysis of pesticide level to control toxicity and for environment conservation. The present review summarises AChE based biosensors by discussing their characteristic features in terms of fabrication, detection limit, linearity range, time of incubation, and storage stability. Use of nanoparticles in recently reported fabrication strategies has improved the efficiency of biosensors to a great extent making them more reliable and robust. PMID:24383001

  8. Chiral analysis of amino acids using electrochemical composite bienzyme biosensors.

    PubMed

    Domínguez, R; Serra, B; Reviejo, A J; Pingarrón, J M

    2001-11-15

    The construction and performance of bienzyme amperometric composite biosensors for the selective determination of l- or d-amino acids is reported. D- or L-Amino acid oxidase, horseradish peroxidase, and the mediator ferrocene were coimmobilized by simple physical inclusion into the bulk of a graphite-70% Teflon electrode matrix. Working conditions including amino acid oxidase loading and pH were optimized. Studies on the repeatability of the amperometric response obtained at +0.00 V, with and without regeneration of the electrode surface by polishing, on the useful lifetime of one single biosensor and on the reproducibility in the fabrication of different biosensors illustrate the robustness of the bioelectrodes design. Calibration plots by both amperometry in stirred solutions and flow injection with amperometric detection were obtained for L-arginine, L-phenylalanine, L-leucine, L-methionine, L-tryptophan, D-leucine, D-methionine, D-serine, and D-valine. Differences in sensitivity were discussed in terms of the hydrophobicity of the substrate and of the electrode surface. The bienzyme composite electrode was applied to the determination of L- and D-amino acids in racemic samples, as well as to the estimation of the L-amino acids content in muscatel grapes. PMID:11700983

  9. Effect of DNA type on response of DNA biosensor for carcinogens

    NASA Astrophysics Data System (ADS)

    Sani, Nor Diyana bt. Md.; Heng, Lee Yook; Surif, Salmijah; Lazim, Azwani Mat

    2013-11-01

    Carcinogens are cancer causing chemicals that can bind to DNA and cause damage to the DNA. These chemicals are available everywhere including in water, air, soil and food. Therefore, a sensor that can detect the presence of these chemicals will be a very useful tool. Since carcinogens bind to DNA, DNA can be used as the biological element in a biosensor. This study has utilized different types of DNA in a biosensor for carcinogen detection. The DNAs include double stranded calf thymus DNA, single stranded calf thymus DNA and guanine rich single stranded DNA. The modified SPE was exposed to a carcinogen followed by interaction with methylene blue which acts as the electroactive indicator. The SPE was then analysed using differential pulse voltammetry (DPV). Optimization studies were conducted for MB concentration and accumulation time, DNA concentration, as well as effect of buffer concentration, buffer pH and ionic strength. The performance of the biosensor was tested on a group 1 carcinogen, formaldehyde. The results indicated that the usage of guanine rich single stranded DNA also gives higher response as carcinogens prefer to bind with guanine compared to other bases.

  10. Biosensors.

    ERIC Educational Resources Information Center

    Rechnitz, Garry A.

    1988-01-01

    Describes theory and principles behind biosensors that incorporate biological components as part of a sensor or probe. Projects major applications in medicine and veterinary medicine, biotechnology, food and agriculture, environmental studies, and the military. Surveys current use of biosensors. (ML)

  11. Multi-channel PMMA microfluidic biosensor with integrated IDUAs for electrochemical detection

    PubMed Central

    Wongkaew, Nongnoot; He, Peng; Kurth, Vanessa; Surareungchai, Werasak; Baeumner, Antje J.

    2013-01-01

    A novel multi-channel poly(methyl methacrylate) (PMMA) microfluidic biosensor with interdigitated ultramicroelectrode arrays (IDUAs) for electrochemical detection was developed. The focus of the development was a simple fabrication procedure and the realization of a reliable large IDUA that can provide detection simultaneously to several microchannels. As proof of concept, five microchannels are positioned over a large single IDUA where the channels are parallel with the length of electrode finger. The IDUAs were fabricated on the PMMA cover piece and bonded to a PMMA substrate containing the microfluidic channels using UV/ozone-assisted thermal bonding. Conditions of device fabrication were optimized realizing a rugged large IDUA within a bonded PMMA device. Gold adhesion to the PMMA, protective coatings and pressure during bonding were optimized. Its electrochemical performance was studied using amperometric detection of potassium ferri and ferro hexacyanide. Cumulative signals within the same chip showed very good linearity over a range of 0 - 38 μM (R2 = 0.98) and a limit of detection of 3.48 μM. The bonding of the device was optimized so that no cross-talk between the channels was observed which otherwise would have resulted in unreliable electrochemical responses. The highly reproducible signals achieved were comparable to those obtained with separate single-channel devices. Subsequently, the multi-channel microfluidic chip was applied to a model bioanalytical detection strategy, i.e. the quantification of specific nucleic acid sequences using a sandwich approach. Here probe-coated paramagnetic beads and probe-tagged liposomes entrapping ferri/ferro hexacyanide as the redox marker were used to bind to a single stranded DNA sequence. Flow rates of the non-ionic detergent n-octyl-β-D-glucopyranoside (OG) for liposome lysis were optimized and the detection of the target sequences was carried out coulometrically within 250 s and with a limit of detection of 12

  12. Simple and rapid fabrication of disposable carbon-based electrochemical cells using an electronic craft cutter for sensor and biosensor applications.

    PubMed

    Afonso, André S; Uliana, Carolina V; Martucci, Diego H; Faria, Ronaldo C

    2016-01-01

    This work describes the construction of an all-plastic disposable carbon-based electrochemical cell (DCell) using a simple procedure based on the use of a home cutter printer for prototyping and laminating. The cutter printer and adhesive vinyl films were used to produce three electrodes in an electrochemical cell layout, and a laminating process was then used to define the geometric area and insulate the electrodes. The DCell showed excellent performance in several applications including the determination of toxic metals in water samples, the immobilization of DNA and the detection of Salmonella. An unmodified DCell was applied for Pb and Cd detection in the range of 100-300 ng mL(-1) with a limit of detection of 50 and 39 ng mL(-1) for Cd and Pb, respectively. DNA was successfully immobilized on a DCell and used for studies of interaction between bisphenol A and DNA. The square wave voltammetry of a DNA modified DCell presented a guanine oxidation current 2.5 times greater after exposure of the electrode to bisphenol A and no current variation for the adenine moiety indicating that bisphenol A showed a preference for DNA interaction sites. A magneto-immunoassay was developed using a DCell for Salmonella detection in milk samples. The system presented a linear range from 100 to 700 cells mL(-1) with a limit of detection of 100 cells mL(-1) and good recovery values between 93% and 101% in milk samples, with no interference from Escherichia coli. Using the proposed method, hundreds of DCells can be assembled in less than two hours, at a material cost of less than US $0.02 per cell. The all-plastic disposable electrochemical cell developed was successfully applied as an electrochemical sensor and biosensor. The feasibility of the developed all-plastic disposable electrochemical cell was demonstrated in applications as both sensor and biosensor. PMID:26695279

  13. Single strand DNA functionalized single wall carbon nanotubes as sensitive electrochemical labels for arsenite detection.

    PubMed

    Wang, Yonghong; Wang, Ping; Wang, Yiqiang; He, Xiaoxiao; Wang, Kemin

    2015-08-15

    In this work, a simple and sensitive electrochemical strategy for arsenite detection based on the ability of arsenite bound to single-strand DNA (ssDNA) and the signal transduction of single wall carbon nanotubes (SWCNTs) is developed. To realize this purpose, the ssDNA/SWCNTs complexes were formed at first by making ssDNA wrapped around SWCNTs via π-stacking. In the presence of arsenite, the arsenite could strongly bind with the G/T bases of ssDNA and decrease the π-π interaction between ssDNA and SWCNTs, resulting in a certain amount of ssDNA dissociating from the complexes. The separated SWCNTs were selectively assembled on the self-assembled monolayer (SAM) modified Au electrode. Then the SWCNTs onto the SAM-modified Au electrode substantially restored heterogeneous electron transfer that was almost totally blocked by the SAM. The assembled SWCNTs could generate a considerably sensitive and specific tactic for signal transduction, which was related to the concentration of the arsenite. Through detecting the currents mediated by SWCNTs, a linear response to concentration of arsenite ranging from 0.5 to 10ppb and a detection limit of 0.5ppb was readily achieved with desirable specificity and sensitivity. Such a SWCNTs-based biosensor creates a simple, sensitive, nonradioactive route for detection of arsenite. In addition, this demonstration provides a new approach to fabrication of stable biosensors with favorable electrochemical properties believed to be appealing to electroanalytical applications. PMID:25966391

  14. Photonic Crystal Biosensor with In-Situ Synthesized DNA Probes for Enhanced Sensitivity

    SciTech Connect

    Hu, Shuren; Zhao, Y.; Retterer, Scott T; Kravchenko, Ivan I; Weiss, Sharon

    2013-01-01

    We report on a nearly 8-fold increase in multi-hole defect photonic crystal biosensor response by incorporating in-situ synthesis of DNA probes, as compared to the conventional functionalization method employing pre-synthesized DNA probe immobilization.

  15. Electrochemical sensors and biosensors based on redox polymer/carbon nanotube modified electrodes: a review.

    PubMed

    Barsan, Madalina M; Ghica, M Emilia; Brett, Christopher M A

    2015-06-30

    The aim of this review is to present the contributions to the development of electrochemical sensors and biosensors based on polyphenazine or polytriphenylmethane redox polymers together with carbon nanotubes (CNT) during recent years. Phenazine polymers have been widely used in analytical applications due to their inherent charge transport properties and electrocatalytic effects. At the same time, since the first report on a CNT-based sensor, their application in the electroanalytical chemistry field has demonstrated that the unique structure and properties of CNT are ideal for the design of electrochemical (bio)sensors. We describe here that the specific combination of phenazine/triphenylmethane polymers with CNT leads to an improved performance of the resulting sensing devices, because of their complementary electrical, electrochemical and mechanical properties, and also due to synergistic effects. The preparation of polymer/CNT modified electrodes will be presented together with their electrochemical and surface characterization, with emphasis on the contribution of each component on the overall properties of the modified electrodes. Their importance in analytical chemistry is demonstrated by the numerous applications based on polymer/CNT-driven electrocatalytic effects, and their analytical performance as (bio) sensors is discussed. PMID:26041516

  16. An impedance-based integrated biosensor for suspended DNA characterization

    PubMed Central

    Ma, Hanbin; Wallbank, Richard W. R.; Chaji, Reza; Li, Jiahao; Suzuki, Yuji; Jiggins, Chris; Nathan, Arokia

    2013-01-01

    Herein, we describe a novel integrated biosensor for performing dielectric spectroscopy to analyze biological samples. We analyzed biomolecule samples with different concentrations and demonstrated that the solution's impedance is highly correlated with the concentration, indicating that it may be possible to use this sensor as a concentration sensor. In contrast with standard spectrophotometers, this sensor offers a low-cost and purely electrical solution for the quantitative analysis of biomolecule solutions. In addition to determining concentrations, we found that the sample solution impedance is highly correlated with the length of the DNA fragments, indicating that the sizes of PCR products could be validated with an integrated chip-based, sample-friendly system within a few minutes. The system could be the basis of a rapid, low-cost platform for DNA characterization with broad applications in cancer and genetic disease research. PMID:24060937

  17. Graphene-metallic nanocomposites as modifiers in electrochemical glucose biosensor transducers

    NASA Astrophysics Data System (ADS)

    Altuntas, Derya Bal; Tepeli, Yudum; Anik, Ulku

    2016-09-01

    Graphene sheets and three different graphene-metallic nanocomposites including graphene-copper (graphene-Cu), graphene-nickel (graphene-Ni) and graphene-platinum (graphene-Pt) were prepared and characterized in the first place. Then the electrochemical performances of these nanocomposites were tested in glucose biosensor transducers, which were formed by combining these metallic nanocomposites with glucose oxidase enzyme and glassy carbon paste electrode (GCPE). This is the first work that includes the usage of these graphene-Me nanocomposites as a part of glucose biosensor transducer. Fabricated amperometric biosensors linear ranges were obtained as follow: For the plain graphene, the linear range was found in the concentration range between 50 μM and 800 μM with the RSD (n = 3 for 50 μM glucose) value of 12.86% and LOD value of 7.2 μM. For graphene-Pt modified glucose biosensor, the linear range was between 10 μM and 600 μM with the RSD (n = 3 for 50 μM glucose) value of 3.45% and LOD value of 3.06 μM. In the case of graphene-Ni modified glucose biosensor, the values were 25 μM to 600 μM with the RSD (n = 3 for 50 μM glucose) value of 8.76% and LOD value of 24.71 μM and for graphene-Cu modified glucose biosensor linear range was 25 μM to 400 μM with the RSD (n = 3 for 50 μM glucose) value of 3.93% and LOD value of 2.87 μM.

  18. An electrochemical sulfite biosensor based on gold coated magnetic nanoparticles modified gold electrode.

    PubMed

    Rawal, Rachna; Chawla, Sheetal; Pundir, Chandra Shekhar

    2012-01-15

    A sulfite oxidase (SO(X)) (EC 1.8.3.1) purified from Syzygium cumini leaves was immobilized onto carboxylated gold coated magnetic nanoparticles (Fe(3)O(4)@GNPs) electrodeposited onto the surface of a gold (Au) electrode through N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC)-N-hydroxy succinimide (NHS) chemistry. An amperometric sulfite biosensor was fabricated using SO(X)/Fe(3)O(4)@GNPs/Au electrode as working electrode, Ag/AgCl as standard and Pt wire as auxiliary electrode. The working electrode was characterized by Fourier Transform Infrared (FTIR) Spectroscopy, Cyclic Voltammetry (CV), Scanning Electron Microscopy (SEM) and Electrochemical Impedance Spectroscopy (EIS) before and after immobilization of SO(X). The biosensor showed optimum response within 2s when operated at 0.2V (vs. Ag/AgCl) in 0.1 M Tris-HCl buffer, pH 8.5 and at 35 °C. Linear range and detection limit were 0.50-1000 μM and 0.15 μM (S/N=3) respectively. Biosensor was evaluated with 96.46% recovery of added sulfite in red wine and 1.7% and 3.3% within and between batch coefficients of variation respectively. Biosensor measured sulfite level in red and white wines. There was good correlation (r=0.99) between red wines sulfite value by standard DTNB (5,5'-dithio-bis-(2-nitrobenzoic acid)) method and the present method. Enzyme electrode was used 300 times over a period of 4 months, when stored at 4 °C. Biosensor has advantages over earlier biosensors that it has excellent electrocatalysis towards sulfite, lower detection limit, higher storage stability and no interference by ascorbate, cysteine, fructose and ethanol. PMID:22035973

  19. Multi-nanomaterial electrochemical biosensor based on label-free graphene for detecting cancer biomarkers.

    PubMed

    Jin, Bing; Wang, Ping; Mao, Hongju; Hu, Bing; Zhang, Honglian; Cheng, Zule; Wu, Zhenhua; Bian, Xiaojun; Jia, Chunping; Jing, Fengxiang; Jin, Qinghui; Zhao, Jianlong

    2014-05-15

    Developing a rapid, accurate and sensitive electrochemical biosensor for detecting cancer biomarkers is important for early detection and diagnosis. This work reports an electrochemical biosensor based on a graphene (GR) platform which is made by CVD, combined with magnetic beads (MBs) and enzyme-labeled antibody-gold nanoparticle bioconjugate. MBs coated with capture antibodies (Ab1) were attached to GR sheets by an external magnetic field, to avoid reducing the conductivity of graphene. Sensitivity was also enhanced by modifying the gold nanoparticles (AuNPs) with horseradish peroxidase (HRP) and the detection antibody (Ab2), to form the conjugate Ab2-AuNPs-HRP. Electron transport between the electrode and analyte target was accelerated by the multi-nanomaterial, and the limit of detection (LOD) for carcinoembryonic antigen (CEA) reached 5 ng mL(-1). The multi-nanomaterial electrode GR/MBs-Ab1/CEA/Ab2-AuNPs-HRP can be used to detect biomolecules such as CEA. The EC biosensor is sensitive and specific, and has potential in the detection of disease markers. PMID:24462797

  20. Gold and Magnetic Nanoparticles-Based Electrochemical Biosensors for Cancer Biomarker Determination.

    PubMed

    Ravalli, Andrea; Marrazza, Giovanna

    2015-05-01

    Detecting cancer at early stage is one of the most important factors associated with the increase of the survival rate of the patients. Cancer biomarkers are able to detect a specific disease early and help to provide treatments before it becomes incurable in later stages. Biomarkers can also be used to determine the recurrence of the disease and to evaluate the follow-up of the patients after a chemio- or radio-therapy and surgery treatments. Electrochemical biosensors are successfully applied for the detection of cancer biomarkers due to their high sensibility, rapid response and low cost. In recent years, the advance in nanotechnology has led to the discovery and the employment of a great number of new materials in nanoscale dimensions. Due to their particular properties, the development of nanostructured biosensors (in particular using gold and magnetic nanoparticles) with high analytical performances increases constantly. In this review recent different strategies for the development of gold and magnetic nanoparticles-based electrochemical biosensors for cancer biomarkers detection were presented. PMID:26504948

  1. Nanoporous gold supported cobalt oxide microelectrodes as high-performance electrochemical biosensors.

    PubMed

    Lang, Xing-You; Fu, Hong-Ying; Hou, Chao; Han, Gao-Feng; Yang, Ping; Liu, Yong-Bing; Jiang, Qing

    2013-01-01

    Tremendous demands for electrochemical biosensors with high sensitivity and reliability, fast response and excellent selectivity have stimulated intensive research on developing versatile materials with ultrahigh electrocatalytic activity. Here we report flexible and self-supported microelectrodes with a seamless solid/nanoporous gold/cobalt oxide hybrid structure for electrochemical nonenzymatic glucose biosensors. As a result of synergistic electrocatalytic activity of the gold skeleton and cobalt oxide nanoparticles towards glucose oxidation, amperometric glucose biosensors based on the hybrid microelectrodes exhibit multi-linear detection ranges with ultrahigh sensitivities at a low potential of 0.26 V (versus Ag/AgCl). The sensitivity up to 12.5 mA mM⁻¹ cm⁻² with a short response time of less than 1 s gives rise to ultralow detection limit of 5 nM. The outstanding performance originates from a novel nanoarchitecture in which the cobalt oxide nanoparticles are incorporated into pore channels of the seamless solid/nanoporous Au microwires, providing excellent electronic/ionic conductivity and mass transport for the enhanced electrocatalysis. PMID:23851924

  2. Synthesis of Electrochemically Reduced Graphene Oxide Bonded to Thiodiazole-Pd and Applications to Biosensor.

    PubMed

    You, Jung-Min; Han, Hyoung Soon; Jeon, Seungwon

    2015-08-01

    A novel biosensor for the determination of hydrogen peroxide and glucose was developed based on EGN-TDZ-Pd, as an electrocatalyst. The preparation of graphene oxide (GO) nanosheets was functionalized by combining it with 5-amino-1,3,4-thiadiazole-2-thiol (TDZ) and by covalently bonding it to palladium (Pd) nanoparticles (GO-TDZ-Pd). In the electrochemical investigation, EGN-TDZ-Pd was characterized via scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and electrochemical impedance spectroscopy (EIS). Cyclic voltammetry (CV) and chronoamperometry (CA) were used to characterize the performance of EGN-TDZ-Pd. The proposed H2O2 biosensor exhibited a wide linear range from 10 µM to 6.5 mM. Also, a glucose biosensor was prepared using glucose oxidase and EGN-TDZ-Pd placed onto a glassy carbon electrode (GCE). The GOx/EGN-TDZ-Pd/GCE was easily prepared using a rapid and simple procedure, and it was utilized for highly sensitive glucose determination. PMID:26369140

  3. An electrochemical biosensor for sensitive detection of microRNA-155: combining target recycling with cascade catalysis for signal amplification.

    PubMed

    Wu, Xiaoyan; Chai, Yaqin; Zhang, Pu; Yuan, Ruo

    2015-01-14

    In this work, a new electrochemical biosensor based on catalyzed hairpin assembly target recycling and cascade electrocatalysis (cytochrome c (Cyt c) and alcohol oxidase (AOx)) for signal amplification was constructed for highly sensitive detection of microRNA (miRNA). It is worth pointing out that target recycling was achieved only based on strand displacement process without the help of nuclease. Moreover, porous TiO2 nanosphere was synthesized, which could offer more surface area for Pt nanoparticles (PtNPs) enwrapping and enhance the amount of immobilized DNA strand 1 (S1) and Cyt c accordingly. With the mimicking sandwich-type reaction, the cascade catalysis amplification strategy was carried out by AOx catalyzing ethanol to acetaldehyde with the concomitant formation of high concentration of H2O2, which was further electrocatalyzed by PtNPs and Cyt c. This newly designed biosensor provided a sensitive detection of miRNA-155 from 0.8 fM to 1 nM with a relatively low detection limit of 0.35 fM. PMID:25495913

  4. Comparison of a prototype magnetoresistive biosensor to standard fluorescent DNA detection.

    PubMed

    Schotter, J; Kamp, P B; Becker, A; Pühler, A; Reiss, G; Brückl, H

    2004-05-15

    We present a comparative analysis of a magnetoresistive biosensor to standard fluorescent DNA detection. The biosensor consists of giant magnetoresistive (GMR) type Cu/Ni(80)Fe(20) multilayers in the second antiferromagnetic coupling maximum. Each of the 206 elements of the magnetoresistive biosensor is patterned into a spiral-shaped line that can cover the area of a typical DNA spot (70 microm diameter). The probe DNA is assembled on top of the sensor elements in different concentrations ranging from 16 pg/microl to 10 ng/microl. Complementary biotin-labeled analyte DNA is hybridized to the probe DNA at a concentration of 10 ng/microl. A number of different commercially available magnetic microspheres are investigated to determine the most appropriate markers. The experimental comparison shows that the relative sensitivity of the magnetoresistive biosensor is superior to the fluorescent detection at low probe DNA concentrations. PMID:15046745

  5. Carbon Nanomaterials Based Electrochemical Sensors/Biosensors for the Sensitive Detection of Pharmaceutical and Biological Compounds.

    PubMed

    Adhikari, Bal-Ram; Govindhan, Maduraiveeran; Chen, Aicheng

    2015-01-01

    Electrochemical sensors and biosensors have attracted considerable attention for the sensitive detection of a variety of biological and pharmaceutical compounds. Since the discovery of carbon-based nanomaterials, including carbon nanotubes, C60 and graphene, they have garnered tremendous interest for their potential in the design of high-performance electrochemical sensor platforms due to their exceptional thermal, mechanical, electronic, and catalytic properties. Carbon nanomaterial-based electrochemical sensors have been employed for the detection of various analytes with rapid electron transfer kinetics. This feature article focuses on the recent design and use of carbon nanomaterials, primarily single-walled carbon nanotubes (SWCNTs), reduced graphene oxide (rGO), SWCNTs-rGO, Au nanoparticle-rGO nanocomposites, and buckypaper as sensing materials for the electrochemical detection of some representative biological and pharmaceutical compounds such as methylglyoxal, acetaminophen, valacyclovir, β-nicotinamide adenine dinucleotide hydrate (NADH), and glucose. Furthermore, the electrochemical performance of SWCNTs, rGO, and SWCNT-rGO for the detection of acetaminophen and valacyclovir was comparatively studied, revealing that SWCNT-rGO nanocomposites possess excellent electrocatalytic activity in comparison to individual SWCNT and rGO platforms. The sensitive, reliable and rapid analysis of critical disease biomarkers and globally emerging pharmaceutical compounds at carbon nanomaterials based electrochemical sensor platforms may enable an extensive range of applications in preemptive medical diagnostics. PMID:26404304

  6. Carbon Nanomaterials Based Electrochemical Sensors/Biosensors for the Sensitive Detection of Pharmaceutical and Biological Compounds

    PubMed Central

    Adhikari, Bal-Ram; Govindhan, Maduraiveeran; Chen, Aicheng

    2015-01-01

    Electrochemical sensors and biosensors have attracted considerable attention for the sensitive detection of a variety of biological and pharmaceutical compounds. Since the discovery of carbon-based nanomaterials, including carbon nanotubes, C60 and graphene, they have garnered tremendous interest for their potential in the design of high-performance electrochemical sensor platforms due to their exceptional thermal, mechanical, electronic, and catalytic properties. Carbon nanomaterial-based electrochemical sensors have been employed for the detection of various analytes with rapid electron transfer kinetics. This feature article focuses on the recent design and use of carbon nanomaterials, primarily single-walled carbon nanotubes (SWCNTs), reduced graphene oxide (rGO), SWCNTs-rGO, Au nanoparticle-rGO nanocomposites, and buckypaper as sensing materials for the electrochemical detection of some representative biological and pharmaceutical compounds such as methylglyoxal, acetaminophen, valacyclovir, β-nicotinamide adenine dinucleotide hydrate (NADH), and glucose. Furthermore, the electrochemical performance of SWCNTs, rGO, and SWCNT-rGO for the detection of acetaminophen and valacyclovir was comparatively studied, revealing that SWCNT-rGO nanocomposites possess excellent electrocatalytic activity in comparison to individual SWCNT and rGO platforms. The sensitive, reliable and rapid analysis of critical disease biomarkers and globally emerging pharmaceutical compounds at carbon nanomaterials based electrochemical sensor platforms may enable an extensive range of applications in preemptive medical diagnostics. PMID:26404304

  7. Carbon nanomaterial-based electrochemical biosensors for label-free sensing of environmental pollutants.

    PubMed

    Ramnani, Pankaj; Saucedo, Nuvia M; Mulchandani, Ashok

    2016-01-01

    Carbon allotropes such as graphene and carbon nanotubes, have been incorporated in electrochemical biosensors for highly sensitive and selective detection of various analytes. The superior physical and electrical properties like high carrier mobility, ambipolar electric field effect, high surface area, flexibility and their compatibility with microfabrication techniques makes these carbon nanomaterials easy to integrate in field-effect transistor (FET)/chemiresistor type configuration which is suitable for portable and point-of-use/field-deployable sensors. This review covers the synthesis of carbon nanostructures (graphene and CNTs) and their integration into devices using various fabrication methods. Finally, we discuss the recent reports showing different sensing platforms that incorporate biomolecules like enzymes, antibodies and aptamers as recognition elements for fabrication of simple, low cost, compact biosensors that can be used for on-site, rapid environmental monitoring of environmental pollutants like pathogens, heavy metals, pesticides and explosives. PMID:25956023

  8. Electrochemical deoxyribonucleic acid biosensor based on the self-assembly film with nanogold decorated on ionic liquid modified carbon paste electrode.

    PubMed

    Gao, Hongwei; Qi, Xiaowei; Chen, Ying; Sun, Wei

    2011-10-17

    An electrochemical DNA biosensor was fabricated by self-assembling probe single-stranded DNA (ssDNA) with a nanogold decorated on ionic liquid modified carbon paste electrode (IL-CPE). IL-CPE was fabricated using 1-butylpyridinium hexafluorophosphate as the binder and the gold nanoparticles were electrodeposited on the surface of IL-CPE (Au/IL-CPE). Then mercaptoacetic acid was self-assembled on the Au/IL-CPE to obtain a layer of modified film, and the ssDNA probe was further covalently-linked with mercaptoacetic acid by the formation of carboxylate ester with the help of N-(3-dimethylamino-propyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide. The hybridization reaction with the target ssDNA was monitored with methylene blue (MB) as the electrochemical indicator. Under the optimal conditions, differential pulse voltammetric responses of MB was proportional to the specific ssDNA arachis sequences in the concentration range from 1.0×10(-11) to 1.0×10(-6) mol L(-1) with the detection limit as 1.5×10(-12) mol L(-1) (3σ). This electrochemical DNA sensor exhibited good stability and selectivity with the discrimination ability of the one-base and three-base mismatched ssDNA sequences. The polymerase chain reaction product of arachis Arabinose operon D gene was successfully detected by the proposed method, which indicated that the electrochemical DNA sensor designed in this paper could be further used for the detection of specific ssDNA sequence. PMID:21907030

  9. Effect of nanoporous gold thin film morphology on electrochemical DNA sensing.

    PubMed

    Daggumati, Pallavi; Matharu, Zimple; Seker, Erkin

    2015-08-18

    Advances in materials science and chemistry have led to the development of a wide range of nanostructured materials for building novel electrochemical biosensors. A systematic understanding of the challenges related to electrode morphology involved in designing such sensors is essential for developing effective biosensing tools. In this study, we use nanoporous gold (np-Au) thin film electrode coatings with submicrometer thicknesses, as a model system to investigate the influence of nanostructuring on DNA-methylene blue (MB) interactions and their application to DNA biosensors. The interaction of single- and double-stranded DNA immobilized onto morphologically different np-Au films with MB was electrochemically interrogated via square wave voltammetry (SWV). The electrochemical signal from these electrodes in response to MB decayed progressively with each SWV scan. The decay rate was governed by accessibility of the electrochemically active np-Au surface by the analyte. The optimum frequency for extracting the maximum signal via SWV was influenced by the film morphology, where the optimum frequency was lower for the nanoporous morphology with lower density of molecular access points into the porous coating. Overall, the np-Au electrodes exhibited a 10-fold enhancement in probe grafting density and approximately 10-fold higher electrochemical current upon probe-target hybridization as compared to the planar Au electrodes. The np-Au electrodes enabled sensitive detection with a dynamic range of 10 to 100 nM that shifts by 1 order of magnitude for coarsened np-Au morphology due to increased target penetration into the porous network and hence enhanced hybridization efficiency. These findings provide insight into the influence of nanostructuring on the transport mechanisms of small molecules and nucleic acids, and yield an understanding of diverse sensor performance parameters such as DNA grafting density, hybridization efficiency, sensitivity and dynamic range. PMID

  10. An ultrasensitive DNA biosensor based on covalent immobilization of probe DNA on fern leaf-like α-Fe2O3 and chitosan Hybrid film using terephthalaldehyde as arm-linker.

    PubMed

    Xu, Biyan; Zheng, Delun; Qiu, Weiwei; Gao, Feng; Jiang, Shaoxiong; Wang, Qingxiang

    2015-10-15

    In this work, a novel electrochemical DNA biosensor has been developed based on the hybrid film of fern leaf-like α-Fe2O3 microparticles and chitosan (CS). The fern leaf-like α-Fe2O3 microparticles were synthesized via a facile template-free hydrothermal method, and their morphologies were characterized by X-ray diffraction, energy dispersive spectrometry, scanning electron microscope, and transmission electron microscope. Electrochemical characterization assays revealed that the hybrid film modified electrode had remarkable synergistic effects of the large accessible surface area and high electrical conductivity of semiconductive Fe2O3, and the good film stability of CS. Based on the rich amino groups on CS, the CS-Fe2O3 hybrid film was employed as a functional matrix for probe DNA immobilization using terephthalaldehyde (TPA) as a bifunctional arm-linker. The hybridization capacity of the developed biosensor was evaluated with electrochemical impedance spectroscopy (EIS) using [Fe(CN)6](3-/4-) as the indicating probe. A wide dynamic detection range from 1.0 × 10(-14) to 1.0 × 10(-10)M with ultralow detection limit of 5.6 × 10(-15)M was achieved for the target DNA. The hybridization selectivity experiments further revealed that the biosensor could discriminate fully complementary sequences from one-base mismatched, three-base mismatched, and non-complementary sequences. Moreover, the biosensor showed the advantage of good regeneration ability and reproducibility. PMID:25982725

  11. Novel and simple electrochemical biosensor monitoring attomolar levels of miRNA-155 in breast cancer.

    PubMed

    Cardoso, Ana R; Moreira, Felismina T C; Fernandes, Rúben; Sales, M Goreti F

    2016-06-15

    This work, describes for the first time, a simple biosensing design to yield an ultrasensitive electrochemical biosensor for a cancer biomarker detection, miRNA-155, with linear response down to the attomolar range. MiRNA-155 was selected for being overexpressed in breast cancer. The biosensor was assembled in two stages: (1) the immobilization of the anti-miRNA-155 that was thiol modified on an Au-screen printed electrode (Au-SPE), followed by (2) blocking the areas of non-specific binding with mercaptosuccinic acid. Atomic force microscopy (AFM) and electrochemical techniques including cyclic voltammetry (CV), impedance spectroscopy (EIS) and square wave voltammetry (SWV) confirmed the surface modification of these devices and their ability to hybridize successfully and stably with miRNA-155. The final biosensor provided a sensitive detection of miRNA-155 from 10 aM to 1.0 nM with a low detection limit (LOD) of 5.7 aM in real human serum samples. Good results were obtained in terms of selectivity towards breast cancer antigen CA-15.3 and bovine serum albumin (BSA). Raw fluid extracts from cell-lines of melanoma did not affect the biosensor response (no significant change of the blank), while raw extracts from breast cancer yielded a positive signal against miRNA-155. This simple and sensitive strategy is a promising alternative for simultaneous quantitative analysis of multiple miRNA in physiological fluids for biomedical research and point-of-care (POC) diagnosis. PMID:26901459

  12. New Catalytic DNA Biosensors for Radionuclides and Metal ions

    SciTech Connect

    Lu, Yi

    2002-06-01

    The goals of the project are to develop new catalytic DNA biosensors for simultaneous detection and quantification of bioavailable radionuclides and metal ions, and apply the sensors for on-site, real-time assessment of concentration, speciation and stability of the individual contaminants during and after bioremediation. A negative selection strategy was tested and validated. In vitro selection was shown to yield highly active and specific transition metal ion-dependent catalytic DNA/RNA. A fluorescence resonance energy transfer (FRET) study of in vitro selected DNA demonstrated that the trifluorophore labeled system is a simple and powerful tool in studying complex biomolecules structure and dynamics, and is capable of revealing new sophisticated structural changes. New fluorophore/quenchers in a single fluorosensor yielded improved signal to noise ratio in detection, identification and quantification of metal contaminants. Catalytic DNA fluorescent and colorimetric sensors were shown useful in sensing lead in lake water and in leaded paint. Project results were described in two papers and two patents, and won an international prize.

  13. New Catalytic DNA Biosensors for Radionuclides and Metal ions

    SciTech Connect

    Lu, Yi

    2003-06-01

    The goals of the project are to develop new catalytic DNA biosensors for simultaneous detection and quantification of bioavailable radionuclides and metal ions, and apply the sensors for on-site, real-time assessment of concentration, speciation and stability of the individual contaminants during and after bioremediation. A negative selection strategy was tested and validated. In vitro selection was shown to yield highly active and specific transition metal ion-dependent catalytic DNA/RNA. A fluorescence resonance energy transfer (FRET) study of in vitro selected DNA demonstrated that the trifluorophore labeled system is a simple and powerful tool in studying complex biomolecules structure and dynamics, and is capable of revealing new sophisticated structural changes. New fluorophore/quenchers in a single fluorosensor yielded improved signal to noise ratio in detection, identification and quantification of metal contaminants. Catalytic DNA fluorescent and colorimetric sensors were shown useful in sensing lead in lake water and in leaded paint. Project results were described in two papers and two patents, and won an international prize.

  14. E-DNA sensor of Mycobacterium tuberculosis based on electrochemical assembly of nanomaterials (MWCNTs/PPy/PAMAM).

    PubMed

    Miodek, Anna; Mejri, Nawel; Gomgnimbou, Michel; Sola, Christophe; Korri-Youssoufi, Hafsa

    2015-09-15

    Two-step electrochemical patterning methods have been employed to elaborate composite nanomaterials formed with multiwalled carbon nanotubes (MWCNTs) coated with polypyrrole (PPy) and redox PAMAM dendrimers. The nanomaterial has been demonstrated as a molecular transducer for electrochemical DNA detection. The nanocomposite MWCNTs-PPy has been formed by wrapping the PPy film on MWCNTs during electrochemical polymerization of pyrrole on the gold electrode. The MWCNTs-PPy layer was modified with PAMAM dendrimers of fourth generation (PAMAM G4) with covalent bonding by electro-oxidation method. Ferrocenyl groups were then attached to the surface as a redox marker. The electrochemical properties of the nanomaterial (MWCNTs-PPy-PAMAM-Fc) were studied using both square wave voltammetry and cyclic voltammetry to demonstrate efficient electron transfer. The nanomaterial shows high performance in the electrochemical detection of DNA hybridization leading to a variation in the electrochemical signal of ferrocene with a detection limit of 0.3 fM. Furthermore, the biosensor demonstrates ability for sensing DNA of rpoB gene of Mycobacterium tuberculosis in real PCR samples. Developed biosensor was suitable for detection of sequences with a single nucleotide polymorphism (SNP) T (TCG/TTG), responsible for resistance of M. tuberculosis to rifampicin drug, and discriminating them from wild-type samples without such mutation. This shows potential of such systems for further application in pathogens diagnostic and therapeutic purpose. PMID:26313137

  15. A Novel Sandwich Electrochemical Immunosensor Based on the DNA-Derived Magnetic Nanochain Probes for Alpha-Fetoprotein

    PubMed Central

    Gan, Ning; Jia, Liyong; Zheng, Lei

    2011-01-01

    One novel electrochemical immunosensor was constructed by immobilizing capture antibody of alpha-fetoprotein (AFP Ab1) on a nafion/nanogold-particle modified glassy carbon electrode. With a sandwich immunoassay, one DNA-derived magnetic nanoprobe, simplified as DNA/(ZMPs—HRP-AFP Ab2)n, was employed for the detection of AFP. The fabricated procedure of the proposed biosensor was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The performance and factors influencing the performance of the biosensor were also evaluated. Under optimal conditions, the developed biosensor exhibited a well-defined electrochemical behavior toward the reduction of AFP ranging from 0.01 to 200 ng/mL with a detection limit of 4 pg/mL (S/N = 3). The biosensor was applied to the determination of AFP in serum with satisfactory results. It is important to note that the sandwich nanochainmodified electro-immunosensor provided an alternative substrate for the immobilization of other tumor markers. PMID:22013390

  16. Direct attachment of DNA to semiconducting surfaces for biosensor applications.

    PubMed

    Fahrenkopf, Nicholas M; Shahedipour-Sandvik, Fatemeh; Tokranova, Natalya; Bergkvist, Magnus; Cady, Nathaniel C

    2010-11-01

    In this work we propose a novel method of immobilizing nucleic acids for field effect or high electron mobility transistor-based biosensors. The naturally occurring 5' terminal phosphate group on nucleic acids was used to coordinate with semiconductor and metal oxide surfaces. We demonstrate that DNA can be directly immobilized onto ZrO(2), AlGaN, GaN, and HfO(2) while retaining its ability to hybridize to target sequences with high specificity. By directly immobilizing the probe molecule to the sensor surface, as opposed to conventional crosslinking strategies, the number of steps in device fabrication is reduced. Furthermore, hybridization to target strands occurs closer to the sensor surface, which has the potential to increase device sensitivity by reducing the impact of the Debye screening length. PMID:20869405

  17. Electrochemical deoxyribonucleic acid biosensor based on electrodeposited graphene and nickel oxide nanoparticle modified electrode for the detection of salmonella enteritidis gene sequence.

    PubMed

    Sun, Wei; Wang, Xiuli; Lu, Yongxi; Gong, Shixing; Qi, Xiaowei; Lei, Bingxin; Sun, Zhenfan; Li, Guangjiu

    2015-04-01

    In this paper a new electrochemical DNA biosensor was prepared by using graphene (GR) and nickel oxide (NiO) nanocomposite modified carbon ionic liquid electrode (CILE) as the substrate electrode. GR and NiO nanoparticles were electrodeposited on the CILE surface step-by-step to get the nanocomposite. Due to the strong affinity of NiO with phosphate groups of ssDNA, oligonucleotide probe with a terminal 5'-phosphate group could be attached on the surface of NiO/GR/CILE, which could further hybridize with the target ssDNA sequence. Methylene blue (MB) was used as the electrochemical indicator for monitoring the hybridization reaction. Under the optimal conditions the reduction peak current of MB was proportional to the concentration of salmonella enteritidis gene sequence in the range from 1.0×10(-13) to 1.0×10(-6)molL(-1) with a detection limit as 3.12×10(-14)molL(-1). This electrochemical DNA sensor exhibited good discrimination ability to one-base and three-base mismatched ssDNA sequences, and the polymerase chain reaction amplification product of salmonella enteritidis gene sequences were further detected with satisfactory results. PMID:25686924

  18. Luminescent Iridium(III) Complex Labeled DNA for Graphene Oxide-Based Biosensors.

    PubMed

    Zhao, Qingcheng; Zhou, Yuyang; Li, Yingying; Gu, Wei; Zhang, Qi; Liu, Jian

    2016-02-01

    There has been growing interest in utilizing highly photostable iridium(III) complexes as new luminescent probes for biotechnology and life science. Herein, iridium(III) complex with carboxyl group was synthesized and activated with N-hydroxysuccinimide, followed by tagging to the amino terminate of single-stranded DNA (ssDNA). The Ir-ssDNA probe was further combined with graphene oxide (GO) nanosheets to develop a GO-based biosensor for target ssDNA detection. The quenching efficiency of GO, and the photostability of iridium(III) complex and GO-Ir-ssDNA biosensor, were also investigated. On the basis of the high luminescence quenching efficiency of GO toward iridium(III) complex, the GO-Ir-ssDNA biosensor exhibited minimal background signals, while strong emission was observed when Ir-ssDNA desorbed from GO nanosheets and formed a double helix with the specific target, leading to a high signal-to-background ratio. Moreover, it was found that luminescent intensities of iridium(III) complex and GO-Ir-ssDNA biosensor were around 15 and 3 times higher than those of the traditional carboxyl fluorescein (FAM) dye and the GO-FAM-ssDNA biosensor after UV irradiation, respectively. Our study suggested the sensitive and selective Ir-ssDNA probe was suitable for the development of highly photostable GO-based detection platforms, showing promise for application beyond the OLED (organic light emitting diode) area. PMID:26753824

  19. DNA interaction of [Cu(dmp)(phen-dion)] (dmp=4,7 and 2,9 dimethyl phenanthroline, phen-dion=1,10-phenanthroline-5,6-dion) complexes and DNA-based electrochemical biosensor using chitosan-carbon nanotubes composite film.

    PubMed

    Kashanian, Soheila; Khodaei, Mohammad Mehdi; Roshanfekr, Hamideh; Peyman, Hossein

    2013-10-01

    The interaction of two new water-soluble [Cu(4,7-dmp)(phen-dione)Cl]Cl (1) and [Cu(2,9-dmp)(phen-dione)Cl]Cl (2) which dmp is dimethyl-1,10-phenanthroline and phen-dion represents 1,10-phenanthroline-5,6-dion, with DNA in solution and immobilized DNA on a chitosan-carbon nanotubes composite modified glassy carbon electrode were investigated by cyclic voltammetry and UV-Vis spectroscopy techniques. In solution interactions, spectroscopic and electrochemical evidences indicate outside binding of these complexes. To clarify the binding mode of complexes, it was done competition studies with Hoechst and Neutral red as groove binder and intercalative probes, respectively. All these results indicating that, these two complexes (1) and (2) interact with DNA via groove binding and partially intercalative modes, respectively. The electrochemical characterization experiments showed that the nanocomposite film of chitosan-carbon nanotubes could effectively immobilize DNA and greatly improve the electron-transfer reactions of the electroactive molecules that latter finding is the result of strong interactions between captured DNA and Cu complexes. This result indicates that these complexes could be noble candidates as hybridization indicators in further studies. At the end, these new complexes showed excellent antitumor activity against K562 (human chronic myeloid leukemia) cell lines. PMID:23811151

  20. Sensitivity improvement of a miniaturized label-free electrochemical impedance biosensor by electrode edge effect

    NASA Astrophysics Data System (ADS)

    Kuo, Yi-Ching; Chen, Ching-Sung; Chang, Ku-Ning; Lin, Chih-Ting; Lee, Chih-Kung

    2014-07-01

    Point-of-care (PoC) biosensors continue to gain popularity because of the desire to improve cost performance in today's health care industry. As cardiovascular disease (CVD) remains one of the top three leading causes of death in Asia, a tool that can help to detect CVDs is highly sought after. We present a high-sensitivity PoC biosensor that can be used to detect CVD biomarkers. To meet the requirements of a PoC biosensor, we adopted electrochemical methods as the basis of the detection. A more stable three-electrode configuration was miniaturized and put onto a biochip. To improve the detection sensitivity associated with the reduced size in the biochip, computer simulation was used to investigate several potential effective possibilities. We found that the electrolyte current density on the edge near the working electrode (WE) and counter electrode (CE) was higher. This was verified using an atomic force microscope to measure the surface potential. We then experimented with the configuration by lengthening the edge of the WE and CE without changing the area of the WE and CE and maintained the gap between the two electrodes. We found improved measurement efficiency with our newly developed biochip.

  1. RNA aptamer-based electrochemical biosensor for selective and label-free analysis of dopamine.

    PubMed

    Farjami, Elaheh; Campos, Rui; Nielsen, Jesper S; Gothelf, Kurt V; Kjems, Jørgen; Ferapontova, Elena E

    2013-01-01

    The inherent redox activity of dopamine enables its direct electrochemical in vivo analysis ( Venton , B. J.; Wightman, M. R. Anal. Chem. 2003, 75, 414A). However, dopamine analysis is complicated by the interference from other electrochemically active endogenous compounds present in the brain, including dopamine precursors and metabolites and other neurotransmitters (NT). Here we report an electrochemical RNA aptamer-based biosensor for analysis of dopamine in the presence of other NT. The biosensor exploits a specific binding of dopamine by the RNA aptamer, immobilized at a cysteamine-modified Au electrode, and further electrochemical oxidation of dopamine. Specific recognition of dopamine by the aptamer allowed a selective amperometric detection of dopamine within the physiologically relevant 100 nM to 5 μM range in the presence of competitive concentrations of catechol, epinephrine, norepinephrine, 3,4-dihydroxy-phenylalanine (L-DOPA), 3,4-dihydroxyphenylacetic acid (DOPAC), methyldopamine, and tyramine, which gave negligible signals under conditions of experiments (electroanalysis at 0.185 V vs Ag/AgCl). The interference from ascorbic and uric acids was eliminated by application of a Nafion-coated membrane. The aptasensor response time was <1 s, and the sensitivity of analysis was 62 nA μM(-1) cm(-2). The proposed design of the aptasensor, based on electrostatic interactions between the positively charged cysteamine-modified electrode and the negatively charged aptamer, may be used as a general strategy not to restrict the conformational freedom and binding properties of surface-bound aptamers and, thus, be applicable for the development of other aptasensors. PMID:23210972

  2. Multi-wall carbon nanotubes (MWCNTs)-doped polypyrrole DNA biosensor for label-free detection of genetically modified organisms by QCM and EIS.

    PubMed

    Truong, Thi Ngoc Lien; Tran, Dai Lam; Vu, Thi Hong An; Tran, Vinh Hoang; Duong, Tuan Quang; Dinh, Quang Khieu; Tsukahara, Toshifumi; Lee, Young Hoon; Kim, Jong Seung

    2010-01-15

    In this paper, we describe DNA electrochemical detection for genetically modified organism (GMO) based on multi-wall carbon nanotubes (MWCNTs)-doped polypyrrole (PPy). DNA hybridization is studied by quartz crystal microbalance (QCM) and electrochemical impedance spectroscopy (EIS). An increase in DNA complementary target concentration results in a decrease in the faradic charge transfer resistance (R(ct)) and signifying "signal-on" behavior of MWCNTs-PPy-DNA system. QCM and EIS data indicated that the electroanalytical MWCNTs-PPy films were highly sensitive (as low as 4pM of target can be detected with QCM technique). In principle, this system can be suitable not only for DNA but also for protein biosensor construction. PMID:20006069

  3. Impedimetric DNA-biosensor for the study of anti-cancer action of mitomycin C: comparison between acid and electroreductive activation.

    PubMed

    Ensafi, Ali A; Amini, Maryam; Rezaei, Behzad

    2014-09-15

    An electrochemical protocol is described for direct monitoring of anti-cancer properties of MMC. Using electrochemical impedance spectroscopy, a pretreated pencil graphite electrode (PGE) modified with multiwall carbon nanotubes (MWCNTs) and poly(diallyldimethylmmonium chloride), PDDA, decorated with ds-DNA was employed in this study to identify DNA damages induced by MMC. The change in charge transfer resistance after incubation of the DNA-biosensor in MMC solution for a known time was used as indication of DNA damage. It was found that MMC did not interact with DNA. As MMC does not inherently possess any anti-cancer activity, it is, therefore, necessary to activate it by either of two ways: activation in acidic media or electrochemical activation. Incubation of DNA-modified electrode in activated MMC led to alterations in DNA and changes in its electrochemical properties (which forms the theme of the present study). Acid and electroreductive MMC activations were compared and different adducts were subsequently generated, suggesting that the drug can bind to DNA in more than one way. Impedance spectroscopy was used for the first time as a novel technique for detecting DNA-drug adducts. PMID:24747202

  4. Electrochemical approach of anticancer drugs--DNA interaction.

    PubMed

    Rauf, S; Gooding, J J; Akhtar, K; Ghauri, M A; Rahman, M; Anwar, M A; Khalid, A M

    2005-02-23

    The interaction of drugs with DNA is among the most important aspects of biological studies in drug discovery and pharmaceutical development processes. In recent years there has been a growing interest in the electrochemical investigation of interaction between anticancer drugs and DNA. Observing the pre and post electrochemical signals of DNA or drug interaction provides good evidence for the interaction mechanism to be elucidated. Also this interaction could be used for the quantification of these drugs and for the determination of new drugs targeting DNA. Electrochemical approach can provide new insight into rational drug design and would lead to further understanding of the interaction mechanism between anticancer drugs and DNA. PMID:15708659

  5. Facile synthesis of tetragonal columnar-shaped TiO2 nanorods for the construction of sensitive electrochemical glucose biosensor.

    PubMed

    Yang, Zhanjun; Tang, Yan; Li, Juan; Zhang, Yongcai; Hu, Xiaoya

    2014-04-15

    A tetragonal columnar-shaped TiO2 (TCS-TiO2) nanorods are synthesized via a facile route for the immobilization of glucose oxidase (GOx). A novel electrochemical glucose biosensor is constructed based on the direct electrochemistry of GOx at TCS-TiO2 modified glassy carbon electrode. The fabricated biosensor is characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, electrochemical impedance spectra and cyclic voltammetry. The immobilized enzyme molecules on TCS-TiO2 nanorods retain its native structure and bioactivity and show a surface controlled, quasi-reversible and fast electron transfer process. The TCS-TiO2 nanorods have large surface area and provide a favorable microenvironment for enhancing the electron transfer between enzyme and electrode surface. The constructed glucose biosensor shows wide linear range from 5.0×10(-6) to 1.32×10(-3) M with a high sensitivity of 23.2 mA M(-1) cm(-2). The detection limit is calculated to be 2.0×10(-6) M at signal-to-noise of 3. The proposed glucose biosensor also exhibits excellent selectivity, good reproducibility, and acceptable operational stability. Furthermore, the biosensor can be successfully applied in the detection of glucose in serum sample at the applied potential of -0.50 V. The TCS-TiO2 nanorods provide an efficient and promising platform for the immobilization of proteins and development of excellent biosensors. PMID:24325982

  6. A sensitive, rapid and inexpensive way to assay pesticide toxicity based on electrochemical biosensor.

    PubMed

    Yong, Daming; Liu, Chang; Yu, Dengbin; Dong, Shaojun

    2011-03-15

    We reported a rapid toxicity assay method using electrochemical biosensor for pesticides, Escherichia coli (E. coli) was taken as a model microorganism for test. In this method, we adopted ferricyanide instead of natural electron acceptor O(2), and then microbial oxidation was substantially accelerated. Toxicity assays measured the effect of toxic materials on the metabolic activity of microorganisms. The current signal of ferrocyanide produced from the metabolism was proven to be directly related to the toxicity, which could be amplified by ultramicroelectrode array (UMEA). The ratio of the electrochemical signals, recorded in the presence and absence of toxin, provided an index of inhibition. Accordingly, a direct toxicity assessment (DTA) based on chronoamperometry was proposed to detect the effect of toxic chemicals on microorganisms. 3,5-Dichlorophenol (DCP) was taken as the reference toxicant, its IC50 was estimated to be 8.0mg/L. Three pesticides were examined using this method. IC50 values of 6.5mg/L for Ametryn, 22 mg/L for Fenamiphos and 5.7 mg/L for Endosulfan were determined and in line with EC50 values reported in the literature. Atomic force microscopy (AFM) was also used for morphology characterization of E. coli induced by three pesticides. These results confirmed the present electrochemical method used is reliable. In addition, the electrochemical method is a sensitive, rapid and inexpensive way for toxicity assays of pesticides. PMID:21315890

  7. Label-free DNA biosensor based on resistance change of platinum nanoparticles assemblies.

    PubMed

    Skotadis, Evangelos; Voutyras, Konstantinos; Chatzipetrou, Marianneza; Tsekenis, Georgios; Patsiouras, Lampros; Madianos, Leonidas; Chatzandroulis, Stavros; Zergioti, Ioanna; Tsoukalas, Dimitris

    2016-07-15

    A novel nanoparticle based biosensor for the fast and simple detection of DNA hybridization events is presented. The sensor utilizes hybridized DNA's charge transport properties, combining them with metallic nanoparticle networks that act as nano-gapped electrodes. The DNA hybridization events can be detected by a significant reduction in the sensor's resistance due to the conductive bridging offered by hybridized DNA. By modifying the nanoparticle surface coverage, which can be controlled experimentally being a function of deposition time, and the structural properties of the electrodes, an optimized biosensor for the in situ detection of DNA hybridization events is ultimately fabricated. The fabricated biosensor exhibits a wide response range, covering four orders of magnitude, a limit of detection of 1nM and can detect a single base pair mismatch between probe and complementary DNA. PMID:26995284

  8. New advances in electrochemical biosensors for the detection of toxins: Nanomaterials, magnetic beads and microfluidics systems. A review.

    PubMed

    Reverté, Laia; Prieto-Simón, Beatriz; Campàs, Mònica

    2016-02-18

    The use of nanotechnology in bioanalytical devices has special advantages in the detection of toxins of interest in food safety and environmental applications. The low levels to be detected and the small size of toxins justify the increasing number of publications dealing with electrochemical biosensors, due to their high sensitivity and design versatility. The incorporation of nanomaterials in their development has been exploited to further increase their sensitivity, providing simple and fast devices, with multiplexed capabilities. This paper gives an overview of the electrochemical biosensors that have incorporated carbon and metal nanomaterials in their configurations for the detection of toxins. Biosensing systems based on magnetic beads or integrated into microfluidics systems have also been considered because of their contribution to the development of compact analytical devices. The roles of these materials, the methods used for their incorporation in the biosensor configurations as well as the advantages they provide to the analyses are summarised. PMID:26826685

  9. Electrochemical Biosensors Based on Ferroceneboronic Acid and Its Derivatives: A Review

    PubMed Central

    Wang, Baozhen; Takahashi, Shigehiro; Du, Xiaoyan; Anzai, Jun-ichi

    2014-01-01

    We review recent progress in the development of electrochemical biosensors based on ferroceneboronic acid (FcBA) and ferrocene (Fc)-modified boronic acids. These compounds can be used to construct electrochemical biosensors because they consist of a binding site (i.e., a boronic acid moiety) and an electrochemically active part (i.e., an Fc residue). By taking advantage of the unique properties of FcBA and its derivatives, electrochemical sensors sensitive to sugars, glycated hemoglobin (HbA1c), fluoride (F−) ions, and so forth have been widely studied. FcBA-based sugar sensors rely on the selective binding of FcBA to 1,2- or 1,3-diol residues of sugars through the formation of cyclic boronate ester bonds. The redox properties of FcBA-sugar adduct differ from those of free FcBA, which forms the basis of the electrochemical determination of sugars. Thus, non-enzymatic glucose sensors are now being actively studied using FcBA and Fc-modified boronic acids as redox markers. Using a similar principle, HbA1c can be detected by FcBA-based electrochemical systems because it contains hydrocarbon chains on the polypeptide chain. HbA1c sensors are useful for monitoring blood glucose levels over the preceding 8–12 weeks. In addition, FcBA and Fc-modified boronic acids have been used for the detection of F− ions due to the selective binding of boronic acid to F− ions. F−-ion sensors may be useful alternatives to conventional ion-selective electrodes sensitive to F− ion. Furthermore, FcBA derivatives have been studied to construct lectin; steroids; nucleotides; salicylic acid; and bacteria sensors. One of the limitations of FcBA-based sensors comes from the fact that FcBA derivatives are added in sample solutions as reagents. FcBA derivatives should be immobilized on the surface of electrodes for developing reagentless sensors. PMID:25587421

  10. Electrochemical biosensors based on ferroceneboronic Acid and its derivatives: a review.

    PubMed

    Wang, Baozhen; Takahashi, Shigehiro; Du, Xiaoyan; Anzai, Jun-Ichi

    2014-09-01

    We review recent progress in the development of electrochemical biosensors based on ferroceneboronic acid (FcBA) and ferrocene (Fc)-modified boronic acids. These compounds can be used to construct electrochemical biosensors because they consist of a binding site (i.e., a boronic acid moiety) and an electrochemically active part (i.e., an Fc residue). By taking advantage of the unique properties of FcBA and its derivatives, electrochemical sensors sensitive to sugars, glycated hemoglobin (HbA1c), fluoride (F(-)) ions, and so forth have been widely studied. FcBA-based sugar sensors rely on the selective binding of FcBA to 1,2- or 1,3-diol residues of sugars through the formation of cyclic boronate ester bonds. The redox properties of FcBA-sugar adduct differ from those of free FcBA, which forms the basis of the electrochemical determination of sugars. Thus, non-enzymatic glucose sensors are now being actively studied using FcBA and Fc-modified boronic acids as redox markers. Using a similar principle, HbA1c can be detected by FcBA-based electrochemical systems because it contains hydrocarbon chains on the polypeptide chain. HbA1c sensors are useful for monitoring blood glucose levels over the preceding 8-12 weeks. In addition, FcBA and Fc-modified boronic acids have been used for the detection of F(-) ions due to the selective binding of boronic acid to F(-) ions. F(-)-ion sensors may be useful alternatives to conventional ion-selective electrodes sensitive to F(-) ion. Furthermore, FcBA derivatives have been studied to construct lectin; steroids; nucleotides; salicylic acid; and bacteria sensors. One of the limitations of FcBA-based sensors comes from the fact that FcBA derivatives are added in sample solutions as reagents. FcBA derivatives should be immobilized on the surface of electrodes for developing reagentless sensors. PMID:25587421

  11. A nanoparticle label/immunochromatographic electrochemical biosensor for rapid and sensitive detection of prostate-specific antigen

    SciTech Connect

    Lin, Ying-Ying; Wang, Jun; Liu, Guodong; Wu, Hong; Wai, Chien M.; Lin, Yuehe

    2008-06-15

    We present a nanoparticle (NP) label/immunochromatographic electrochemical biosensor (IEB) for rapid and sensitive detection of prostate-specific antigen (PSA) in human serum. This IEB integrates the immunochromatographic strip with the electrochemical detector for transducing quantitative signals. The NP label, made of CdSe@ZnS, serves as a signal-amplifier vehicle. A sandwich immunoreaction was performed on the immunochromatographic strip. The captured NP labels in the test zone were determined by highly sensitive stripping voltammetric measurement of the dissolved metallic component (cadmium) with a disposable-screen-printed electrode, which is embedded underneath the membrane of the test zone. Experimental parameters (e.g., immunoreaction time, the amount of anti-PSA-NP conjugations applied) and electrochemical detection conditions (e.g., preconcentration potential and time) were optimized using this biosensor for PSA detection. The analytical performance of this biosensor was evaluated with serum PSA samples according to the “figure-of-merits” (e.g., dynamic range, reproducibility, and detection limit). The results were validated with enzyme-linked immunosorbent assay (ELISA) and show high consistency. It is found that this biosensor is very sensitive with the detection limit of 0.02 ng/mL PSA and is quite reproducible. This method is rapid, clinically accurate, and less expensive than other diagnosis tools for PSA; therefore, this IEB coupled with a portable electrochemical analyzer shows great promise for simple, sensitive, quantitative point-of-care testing of disease-related protein biomarkers.

  12. Monitoring of malolactic fermentation in wine using an electrochemical bienzymatic biosensor for L-lactate with long term stability.

    PubMed

    Giménez-Gómez, Pablo; Gutiérrez-Capitán, Manuel; Capdevila, Fina; Puig-Pujol, Anna; Fernández-Sánchez, César; Jiménez-Jorquera, Cecilia

    2016-01-28

    L-lactic acid is monitored during malolactic fermentation process of wine and its evolution is strongly related with the quality of the final product. The analysis of L-lactic acid is carried out off-line in a laboratory. Therefore, there is a clear demand for analytical tools that enabled real-time monitoring of this process in field and biosensors have positioned as a feasible alternative in this regard. The development of an amperometric biosensor for L-lactate determination showing long-term stability is reported in this work. The biosensor architecture includes a thin-film gold electrochemical transducer selectively modified with an enzymatic membrane, based on a three-dimensional matrix of polypyrrole (PPy) entrapping lactate oxidase (LOX) and horseradish peroxidase (HRP) enzymes. The experimental conditions of the biosensor fabrication regarding the pyrrole polymerization and the enzymes entrapment are optimized. The biosensor response to L-lactate is linear in a concentration range of 1 × 10(-6)-1 × 10(-4) M, with a detection limit of 5.2 × 10(-7) M and a sensitivity of - (13500 ± 600) μA M(-1) cm(-2). The biosensor shows an excellent working stability, retaining more than 90% of its original sensitivity after 40 days. This is the determining factor that allowed for the application of this biosensor to monitor the malolactic fermentation of three red wines, showing a good agreement with the standard colorimetric method. PMID:26755146

  13. Rational Design of Bioelectrochemically Multifunctional Film with Oxidase, Ferrocene, and Graphene Oxide for Development of in Vivo Electrochemical Biosensors.

    PubMed

    Wang, Xiuyun; Li, Qian; Xu, Jingjing; Wu, Shuo; Xiao, Tongfang; Hao, Jie; Yu, Ping; Mao, Lanqun

    2016-06-01

    This study demonstrates a new strategy to develop in vivo electrochemical biosensors through rational design and simple formation of bioelectrochemically multifunctional film (BMF). The BMF is rationally designed by first efficiently incorporating oxidase, ferrocene mediator, and graphene oxide into polymaleimidostyrene/polystyrene (PMS/PS) matrix to form a homogeneous mixture and then simply formed by drop-coating the mixture onto solid conducting substrate. By using the as-formed BMF, electrochemical biosensors could be constructed with a technical simplicity and high reproducibility. To illustrate the BMF-based biosensors for in vivo applications, we directly couple the biosensors to in vivo microdialysis to establish an online electrochemical system (OECS) for in vivo monitoring of glucose in rat auditory cortex during salicylate-induced tinnitus model. The OECS with the BMF-based biosensor as the detector shows a linear response toward glucose within a concentration range from 50 to 500 μM with a detection limit of 10 μM (S/N = 3). Additionally, the OECS is stable and does not suffer from the interference from the electroactive species endogenously coexisting in the brain microdialysate. With the BMF-based OECS, the basal level of glucose in the microdialysate continuously sampled from rat auditory cortex is determined to be 120 ± 10 μM (n = 5). After the rats were administrated with salicylate to induce transient tinnitus, the microdialysate glucose concentration in the rat auditory cortex remarkably increased to 433 ± 190 μM (n = 5) at the time point of 1.5 h. This study essentially offers a new, technically simple and reproducible approach to development of in vivo electrochemical biosensors, which is envisaged to be relatively useful for understanding of the molecular basis of brain functions. PMID:27146343

  14. BIOSENSORS

    EPA Science Inventory

    It has recently been proposed under the International Union of Pure and Applied Chemistry (IUPAC) Commission that biosensors be regarded as a subgroup of chemical sensors in which a biologically based mechanism is used for detection of the analyte. hemical sensors are defined und...

  15. An overview of the optical and electrochemical methods for detection of DNA - drug interactions.

    PubMed

    Aleksić, Mara M; Kapetanović, Vera

    2014-01-01

    A large number of inorganic and organic compounds is able to bind to DNA and form complexes. Among them, drugs are very important, especially chemotherapeutics. This paper presents the overview of DNA structural characteristics and types of interactions (covalent and non-covalent) between DNA molecule and drugs. Covalent binding of the drug is irreversible and leads to complete inhibition of DNA function, what conclusively, causes the cell death. On the other hand, non-covalent binding is reversible and based on the principle of molecular recognition. Special attention is given to elucidation of the specific sites in DNA molecule for drug binding. According to their structural characteristics, drugs that react non-covalently with DNA are mainly intercalators, but also minor and major groove binders. When the complex between drug and DNA is formed, both the drug molecule, as well as DNA, experienced some modifications. This paper presents the overview of the methods used for the study of the interactions between DNA and drugs with the aim of detection and explanation of the resulting changes. For this purpose many spectroscopic methods like UV/VIS, fluorescence, infrared and NMR, polarized light spectroscopies like circular and linear dichroism, and fluorescence anisotropy or resonance is used. The development of the electrochemical DNA biosensors has opened a wide perspective using particularly sensitive and selective electrochemical methods for the detection of specific DNA interactions. The presented results summarize literature data obtained by the mentioned methods. The results are used to confirm the DNA damage, to determine drug binding sites and sequence preference, as well as conformational changes due to drug-DNA interaction. PMID:25286211

  16. Combination of cascade chemical reactions with graphene-DNA interaction to develop new strategy for biosensor fabrication.

    PubMed

    Zhu, Xiaoli; Sun, Liya; Chen, Yangyang; Ye, Zonghuang; Shen, Zhongming; Li, Genxi

    2013-09-15

    Graphene, a single atom thick and two dimensional carbon nano-material, has been proven to possess many unique properties, one of which is the recent discovery that it can interact with single-stranded DNA through noncovalent π-π stacking. In this work, we demonstrate that a new strategy to fabricate many kinds of biosensors can be developed by combining this property with cascade chemical reactions. Taking the fabrication of glucose sensor as an example, while the detection target, glucose, may regulate the graphene-DNA interaction through three cascade chemical reactions, electrochemical techniques are employed to detect the target-regulated graphene-DNA interaction. Experimental results show that in a range from 5μM to 20mM, the glucose concentration is in a natural logarithm with the logarithm of the amperometric response, suggesting a best detection limit and detection range. The proposed biosensor also shows favorable selectivity, and it has the advantage of no need for labeling. What is more, by controlling the cascade chemical reactions, detection of a variety of other targets may be achieved, thus the strategy proposed in this work may have a wide application potential in the future. PMID:23542067

  17. Direct ultrasensitive electrochemical biosensing of pathogenic DNA using homogeneous target-initiated transcription amplification

    NASA Astrophysics Data System (ADS)

    Yan, Yurong; Ding, Shijia; Zhao, Dan; Yuan, Rui; Zhang, Yuhong; Cheng, Wei

    2016-01-01

    Sensitive and specific methodologies for detection of pathogenic gene at the point-of-care are still urgent demands in rapid diagnosis of infectious diseases. This work develops a simple and pragmatic electrochemical biosensing strategy for ultrasensitive and specific detection of pathogenic nucleic acids directly by integrating homogeneous target-initiated transcription amplification (HTITA) with interfacial sensing process in single analysis system. The homogeneous recognition and specific binding of target DNA with the designed hairpin probe triggered circular primer extension reaction to form DNA double-strands which contained T7 RNA polymerase promoter and served as templates for in vitro transcription amplification. The HTITA protocol resulted in numerous single-stranded RNA products which could synchronously hybridized with the detection probes and immobilized capture probes for enzyme-amplified electrochemical detection on the biosensor surface. The proposed electrochemical biosensing strategy showed very high sensitivity and selectivity for target DNA with a dynamic response range from 1 fM to 100 pM. Using salmonella as a model, the established strategy was successfully applied to directly detect invA gene from genomic DNA extract. This proposed strategy presented a simple, pragmatic platform toward ultrasensitive nucleic acids detection and would become a versatile and powerful tool for point-of-care pathogen identification.

  18. Direct ultrasensitive electrochemical biosensing of pathogenic DNA using homogeneous target-initiated transcription amplification

    PubMed Central

    Yan, Yurong; Ding, Shijia; Zhao, Dan; Yuan, Rui; Zhang, Yuhong; Cheng, Wei

    2016-01-01

    Sensitive and specific methodologies for detection of pathogenic gene at the point-of-care are still urgent demands in rapid diagnosis of infectious diseases. This work develops a simple and pragmatic electrochemical biosensing strategy for ultrasensitive and specific detection of pathogenic nucleic acids directly by integrating homogeneous target-initiated transcription amplification (HTITA) with interfacial sensing process in single analysis system. The homogeneous recognition and specific binding of target DNA with the designed hairpin probe triggered circular primer extension reaction to form DNA double-strands which contained T7 RNA polymerase promoter and served as templates for in vitro transcription amplification. The HTITA protocol resulted in numerous single-stranded RNA products which could synchronously hybridized with the detection probes and immobilized capture probes for enzyme-amplified electrochemical detection on the biosensor surface. The proposed electrochemical biosensing strategy showed very high sensitivity and selectivity for target DNA with a dynamic response range from 1 fM to 100 pM. Using salmonella as a model, the established strategy was successfully applied to directly detect invA gene from genomic DNA extract. This proposed strategy presented a simple, pragmatic platform toward ultrasensitive nucleic acids detection and would become a versatile and powerful tool for point-of-care pathogen identification. PMID:26729209

  19. Highly sensitive and stable electrochemical sulfite biosensor incorporating a bacterial sulfite dehydrogenase.

    PubMed

    Kalimuthu, Palraj; Tkac, Jan; Kappler, Ulrike; Davis, Jason J; Bernhardt, Paul V

    2010-09-01

    This paper describes a highly sensitive electrochemical (voltammetric) determination of sulfite using a combination of Starkeya novella sulfite dehydrogenase (SDH), horse heart cytochrome c (cyt c), and a self-assembled monolayer of 11-mercaptoundecanol (MU) cast on a gold electrode. The biosensor was optimized in terms of pH and the ratio of cyt c/SDH. The electrocatalytic oxidation current of sulfite increased linearly from 1 to 6 microM at the enzyme-modified electrode with a correlation coefficient of 0.9995 and an apparent Michaelis constant (K(M,app)) of 43 microM. Using an amperometric method, the low detection limit for sulfite at the enzyme-modified electrode was 44 pM (signal-to-noise ratio = 3). The modified electrode retained a stable response for 3 days while losing only ca. 4% of its initial sensitivity during a 2 week storage period in 50 mM Tris buffer solution at 4 degrees C. The enzyme electrode was successfully used for the determination of sulfite in beer and white wine samples. The results of these electrochemical analyses agreed well with an independent spectrophotometric method using Ellman's reagent, but the detection limit was far superior using the electrochemical method. PMID:20698497

  20. A single DNA aptamer functions as a biosensor for ricin.

    PubMed

    Lamont, Elise A; He, Lili; Warriner, Keith; Labuza, Theodore P; Sreevatsan, Srinand

    2011-10-01

    The use of microorganisms or toxins as weapons of death and fear is not a novel concept; however, the modes by which these agents of bioterrorism are deployed are increasingly clever and insidious. One mechanism by which biothreats are readily disseminated is through a nation's food supply. Ricin, a toxin derived from the castor bean plant, displays a strong thermostability and remains active at acidic and alkaline pHs. Therefore, the CDC has assigned ricin as a category B reagent since it may be easily amendable as a deliberate food biocontaminate. Current tools for ricin detection utilize enzymatic activity, immunointeractions and presence of castor bean DNA. Many of these tools are confounded by complex food matrices, display a limited dynamic range of detection and/or lack specificity. Aptamers, short RNA and single stranded DNA sequences, have increased affinity to their selected receptors, experience little cross-reactivity to other homologous compounds and are currently being sought after as biosensors for bacterial contaminants in food. This paper describes the selection and characterization of a single, dominant aptamer, designated as SSRA1, against the B-chain of ricin. SSRA1 displays one folding conformation that is stable across 4-63 °C (ΔG = -5.05). SSRA1 is able to concentrate at least 30 ng mL(-1) of ricin B chain from several liquid food matrices and outcompetes a currently available ELISA kit and ricin aptamer. Furthermore, we show detection of 25 ng mL(-1) of intact ricin A-B complex using SSRA1 combined with surface enhanced Raman scattering technique. Thus, SSRA1 would serve well as pre-analytical tool for processing of ricin from liquid foods to aid current diagnostics as well as a sensor for direct ricin detection. PMID:21748194

  1. A Graphene-Based Biosensing Platform Based on Regulated Release of an Aptameric DNA Biosensor.

    PubMed

    Mao, Yu; Chen, Yongli; Li, Song; Lin, Shuo; Jiang, Yuyang

    2015-01-01

    A novel biosensing platform was developed by integrating an aptamer-based DNA biosensor with graphene oxide (GO) for rapid and facile detection of adenosine triphosphate (ATP, as a model target). The DNA biosensor, which is locked by GO, is designed to contain two sensing modules that include recognition site for ATP and self-replication track that yields the nicking domain for Nt.BbvCI. By taking advantage of the different binding affinity of single-stranded DNA, double-stranded DNA and aptamer-target complex toward GO, the DNA biosensor could be efficiently released from GO in the presence of target with the help of a complementary DNA strand (CPDNA) that partially hybridizes to the DNA biosensor. Then, the polymerization/nicking enzyme synergetic isothermal amplification could be triggered, leading to the synthesis of massive DNA amplicons, thus achieving an enhanced sensitivity with a wide linear dynamic response range of four orders of magnitude and good selectivity. This biosensing strategy expands the applications of GO-DNA nanobiointerfaces in biological sensing, showing great potential in fundamental research and biomedical diagnosis. PMID:26569239

  2. A Graphene-Based Biosensing Platform Based on Regulated Release of an Aptameric DNA Biosensor

    PubMed Central

    Mao, Yu; Chen, Yongli; Li, Song; Lin, Shuo; Jiang, Yuyang

    2015-01-01

    A novel biosensing platform was developed by integrating an aptamer-based DNA biosensor with graphene oxide (GO) for rapid and facile detection of adenosine triphosphate (ATP, as a model target). The DNA biosensor, which is locked by GO, is designed to contain two sensing modules that include recognition site for ATP and self-replication track that yields the nicking domain for Nt.BbvCI. By taking advantage of the different binding affinity of single-stranded DNA, double-stranded DNA and aptamer-target complex toward GO, the DNA biosensor could be efficiently released from GO in the presence of target with the help of a complementary DNA strand (CPDNA) that partially hybridizes to the DNA biosensor. Then, the polymerization/nicking enzyme synergetic isothermal amplification could be triggered, leading to the synthesis of massive DNA amplicons, thus achieving an enhanced sensitivity with a wide linear dynamic response range of four orders of magnitude and good selectivity. This biosensing strategy expands the applications of GO-DNA nanobiointerfaces in biological sensing, showing great potential in fundamental research and biomedical diagnosis. PMID:26569239

  3. Highly sensitive amperometric biosensor based on electrochemically-reduced graphene oxide-chitosan/hemoglobin nanocomposite for nitromethane determination.

    PubMed

    Wen, Yunping; Wen, Wei; Zhang, Xiuhua; Wang, Shengfu

    2016-05-15

    Nitromethane (CH3NO2) is an important organic chemical raw material with a wide variety of applications as well as one of the most common pollutants. Therefore it is pretty important to establish a simple and sensitive detection method for CH3NO2. In our study, a novel amperometric biosensor for nitromethane (CH3NO2) based on immobilization of electrochemically-reduced graphene oxide (rGO), chitosan (CS) and hemoglobin (Hb) on a glassy carbon electrode (GCE) was constructed. Scanning electron microscopy, infrared spectroscopy and electrochemical methods were used to characterize the Hb-CS/rGO-CS composite film. The effects of scan rate and pH of phosphate buffer on the biosensor have been studied in detail and optimized. Due to the graphene and chitosan nanocomposite, the developed biosensor demonstrating direct electrochemistry with faster electron-transfer rate (6.48s(-1)) and excellent catalytic activity towards CH3NO2. Under optimal conditions, the proposed biosensor exhibited fast amperometric response (<5s) to CH3NO2 with a wide linear range of 5 μM~1.46 mM (R=0.999) and a low detection limit of 1.5 μM (S/N=3). In addition, the biosensor had high selectivity, reproducibility and stability, providing the possibility for monitoring CH3NO2 in complex real samples. PMID:26800205

  4. Label-free electrochemical DNA sensor using "click"-functionalized PEDOT electrodes.

    PubMed

    Galán, Teresa; Prieto-Simón, Beatriz; Alvira, Margarita; Eritja, Ramón; Götz, Günther; Bäuerle, Peter; Samitier, Josep

    2015-12-15

    Here we describe a label-free electrochemical DNA sensor based on poly(3,4-ethylenedioxythiophene)-modified (PEDOT-modified) electrodes. An acetylene-terminated DNA probe, complementary to a specific "Hepatitis C" virus sequence, was immobilized onto azido-derivatized conducting PEDOT electrodes using "click" chemistry. DNA hybridization was then detected by differential pulse voltammetry, evaluating the changes in the electrochemical properties of the polymer produced by the recognition event. A limit of detection of 0.13 nM was achieved using this highly selective PEDOT-based genosensor, without the need for labeling techniques or microelectrode fabrication processes. These results are promising for the development of label-free and reagentless DNA hybridization sensors based on conducting polymeric substrates. Biosensors can be easily prepared using any DNA sequence containing an alkyne moiety. The data presented here reveal the potential of this DNA sensor for diagnostic applications in the screening of diseases, such as "Hepatitis C", and genetic mutations. PMID:26210592

  5. Electrochemical performance and biosensor application of TiO2 nanotube arrays with mesoporous structures constructed by chemical etching.

    PubMed

    Wang, Jinwen; Xu, Guangqing; Zhang, Xu; Lv, Jun; Zhang, Xinyi; Zheng, Zhixiang; Wu, Yucheng

    2015-04-28

    Novel mesoporous TiO2 nanotube arrays (TiO2 NTAs) were synthesized by an anodization method combined with chemical etching in HF solution, and the electrochemical performance was studied. Glucose oxidase (GOx) was immobilized on the mesoporous TiO2 NTAs to achieve an efficient biosensor for amperometric detection of glucose. The morphology, structure, component and electrochemical performance of mesoporous TiO2 NTAs were characterized by scanning electron microscopy, high resolution transmission electron microscopy, X-ray diffractometry, X-ray photoelectron spectrometry and an electrochemical workstation, respectively. The influence of the mesoporous structure on the electrochemical performance is discussed in detail by comparing the cyclic voltammograms and electrochemical impedance spectrum of TiO2 and mesoporous TiO2 NTAs in different conditions. High electrochemical active surface area and electron transfer rate play key roles in enhancing the electrochemical performance of mesoporous TiO2 NTAs. When used as the basis of a biosensor, the amperometric response of glucose on a GOx/TiO2-0.5 NTAs electrode is linearly proportion to the glucose concentration in the range from 0.1 to 6 mM with a sensitivity of 0.954 μA mM(-1) cm(-2), which is 14.3 times that of un-etched GOx/TiO2 NTAs. PMID:25811301

  6. On the sensitivity improvement of a miniaturized label-free electrochemical impedance biosensor

    NASA Astrophysics Data System (ADS)

    Kuo, Yi-Ching; Chou, Shin-Ting; Tsai, Pei-I.; Li, Guan-Wei; Lin, Chih-Ting; Lee, Chih-Kung

    2014-03-01

    Development of point-of-care biosensors continues to gain popularity due to the demand of improving the cost performance in today's health care. As cardiovascular disease induced death remains on the top 3 death causes for most Asian countries, this paper is to present a high-sensitivity point-of-care biosensor for the detection of cardiovascular disease biomarkers. To meet the point-of-care biosensors requirements, which include characteristics such as small size, low cost, and ease of operation, we adopted electrochemical methods as the basis of detection. The 4-aminothiophenol was adopted as the bio-linkers to facilitate the antibody-antigen interaction. A more stable three-electrode configuration was miniaturized and laid out onto a biochip. A microfluidics subsystem based on opto-piezoelectronic technology was also integrated to create the microfluidic biochip system. To improve the detection sensitivity associated with the reduction in biochip size, electrochemistry simulation was used to investigate several potentially effective means. We found that the electric field on the edge near working electrode and counter electrode was higher, which was verified by using atomic force microscopy to measure the surface potential. With the successful verification, we explored the configuration, i.e., lengthened the edge of working electrode and counter electrode without changing the areas of working electrode and counter electrode and the gap between these two electrodes, so as to evaluate the possibility of improving the measurement efficiency in our newly developed biochips. Detailed design, simulation and experimental results, improved design identified, etc. were all presented in detail.

  7. Electrochemical DNA sensor-based strategy for sensitive detection of DNA demethylation and DNA demethylase activity.

    PubMed

    Shen, Qingming; Fan, Mengxing; Yang, Yin; Zhang, Hui

    2016-08-31

    DNA demethylation and demethylase activity play important roles in DNA self-repair, and their detection is key to early diagnosis of fatal diseases. Herein, a facile electrochemical DNA (E-DNA) sensor was developed for the sensitive detection of DNA demethylation and demethylase activity based on an enzyme cleavage strategy. The thiol modified hemi-methylated hairpin probe DNA (pDNA) was self-assembled on a Au electrode surface through the formation of AuS bonds. The hemi-methylated pDNA served as the substrate of DNA demethylase (using methyl-CpG-binding domain protein 2 (MBD2) as an example). Following demethylation, the hairpin stem was then recognized and cleaved by BstUI endonuclease. The ferrocene carboxylic acid (FcA)-tagged pDNA strands were released into the buffer solution from the electrode surface, resulting in a significant decrease of electrochemical signal and providing a means to observe DNA demethylation. The activity of DNA demethylase was analyzed in the concentration ranging from 0.5 to 500 ng mL(-1) with a limit of detection as low as 0.17 ng mL(-1). With high specificity and sensitivity, rapid response, and low cost, this simple E-DNA sensor provides a unique platform for the sensitive detection of DNA demethylation, DNA demethylase activity, and related molecular diagnostics and drug screening. PMID:27506345

  8. In-situ detection of DNA hybridization with a microfiber Bragg grating biosensor

    NASA Astrophysics Data System (ADS)

    Sun, Dandan; Guo, Tuan; Xie, Xiaodong; Ran, Yang; Huang, Yunyun; Guan, Bai-Ou

    2014-05-01

    Microfiber Bragg gratings (mFBGs) can be used as cost-effective and relatively simple-to-implement biosensors for monitoring DNA interactions in situ. The sensors are functionalized by a monolayer of poly-L-lysine (PLL) with the specific molecular recognition probe DNA sequences to bind with high specificity to a given target. By recording the wavelength seperation between the two resonant peaks of a single mFBG, the mFBG biosensor is capable of detecting the presence of specific target DNA in situ.

  9. Glycoprofiling of cancer biomarkers: Label-free electrochemical lectin-based biosensors

    PubMed Central

    Pihíková, Dominika; Kasák, Peter

    2016-01-01

    Glycosylation of biomolecules is one of the most prevalent post- and co-translational modification in a human body, with more than half of all human proteins being glycosylated. Malignant transformation of cells influences glycosylation machinery resulting in subtle changes of the glycosylation pattern within the cell populations as a result of cancer. Thus, an altered terminal glycan motif on glycoproteins could provide a warning signal about disease development and progression and could be applied as a reliable biomarker in cancer diagnostics. Among all highly effective glycoprofiling tools, label-free electrochemical impedance spectroscopy (EIS)-based biosensors have emerged as especially suitable tool for point-of-care early-stage cancer detection. Herein, we highlight the current challenges in glycoprofiling of various cancer biomarkers by ultrasensitive impedimetric-based biosensors with low sample consumption, low cost fabrication and simple miniaturization. Additionally, this review provides a short introduction to the field of glycomics and lectinomics and gives a brief overview of glycan alterations in different types of cancer. PMID:27275016

  10. Dynamics of an electrochemical biosensor for the detection of toxic substances in water

    NASA Astrophysics Data System (ADS)

    Simon, Laurent; Ospina, Juan

    2016-05-01

    A proposed analytical method focuses on electrolyte transport to the electrode of an electrochemical cell. The recombinant Escherichia coli whole-cell biosensor detects toxicity in water based on a set of biochemical reactors. Previous contributions elucidated the kinetics of product formation and validated a mathematical model for its diffusion in the chamber. This work introduces an approach to investigate the dynamics of the probe using Laplace transforms and an effective time constant. The transfer function between the electrolyte production and the total current revealed a faster response for larger electrode radii. Both the first-order and effective time constants increased with the chamber height and radius. Separation of variables yields closed-form solutions and helps estimate the kinetics of p-aminophenol generation. When the bacteria were exposed to phenol concentrations of 1.6, 8.3 and 16 ppm, the corresponding overall rate constants were 5.11x10-7, 1.13x10-6 and 1.99x10-6 (product concentration unit/s2), respectively. In addition to parameter estimation, the method can be applied to perform sensitivity analysis and aid manufacturers in meeting design specifications of biosensors.

  11. A novel bi-enzyme electrochemical biosensor for selective and sensitive determination of methyl salicylate.

    PubMed

    Fang, Yi; Umasankar, Yogeswaran; Ramasamy, Ramaraja P

    2016-07-15

    An amperometric sensor based on a bi-enzyme modified electrode was fabricated to detect methyl salicylate, a volatile organic compound released by pathogen-infected plants via systemic response. The detection is based on cascadic conversion reactions that result in an amperometric electrochemical signal. The bi-enzyme electrode is made of alcohol oxidase and horseradish peroxidase enzymes immobilized on to a carbon nanotube matrix through a molecular tethering method. Methyl salicylate undergoes hydrolysis to form methanol, which is consumed by alcohol oxidase to form formaldehyde while simultaneously reducing oxygen to hydrogen peroxide. The hydrogen peroxide will be further reduced to water by horseradish peroxidase, which results in an amperometric signal via direct electron transfer. The bi-enzyme biosensor was evaluated by cyclic voltammetry and constant potential amperometry using hydrolyzed methyl salicylate as the analyte. The sensitivity of the bi-enzyme biosensor as determined by cyclic voltammetry and constant potential amperometry were 112.37 and 282.82μAcm(-2)mM(-1) respectively, and the corresponding limits of detection were 22.95 and 0.98μM respectively. Constant potential amperometry was also used to evaluate durability, repeatability and interference from other compounds. Wintergreen oil was used for real sample study to establish the application of the bi-enzyme sensor for selective determination of plant pathogen infections. PMID:26918616

  12. Electrochemical Glucose Biosensor Based on Glucose Oxidase Displayed on Yeast Surface.

    PubMed

    Wang, Hongwei; Lang, Qiaolin; Liang, Bo; Liu, Aihua

    2015-01-01

    The conventional enzyme-based biosensor requires chemical or physical immobilization of purified enzymes on electrode surface, which often results in loss of enzyme activity and/or fractions immobilized over time. It is also costly. A major advantage of yeast surface display is that it enables the direct utilization of whole cell catalysts with eukaryote-produced proteins being displayed on the cell surface, providing an economic alternative to traditional production of purified enzymes. Herein, we describe the details of the display of glucose oxidase (GOx) on yeast cell surface and its application in the development of electrochemical glucose sensor. In order to achieve a direct electrochemistry of GOx, the entire cell catalyst (yeast-GOx) was immobilized together with multiwalled carbon nanotubes on the electrode, which allowed sensitive and selective glucose detection. PMID:26060079

  13. Novel integrated and portable endotoxin detection system based on an electrochemical biosensor.

    PubMed

    Zuzuarregui, Ana; Souto, David; Pérez-Lorenzo, Eva; Arizti, Fernando; Sánchez-Gómez, Susana; Martínez de Tejada, Guillermo; Brandenburg, Klaus; Arana, Sergio; Mujika, Maite

    2015-01-21

    This paper describes the design, implementation and validation of a sensitive and integral technology solution for endotoxin detection. The unified and portable platform is based on the electrochemical detection of endotoxins using a synthetic peptide immobilized on a thin-film biosensor. The work covers the fabrication of an optimized sensor, the biofunctionalization protocol and the design and implementation of the measuring and signalling elements (a microfluidic chamber and a portable potentiostat-galvanostat), framed ad hoc for this specific application. The use of thin-film technologies to fabricate the biosensing device and the application of simple immobilization and detection methods enable a rapid, easy and sensitive technique for in situ and real time LPS detection. PMID:25431806

  14. Highly selective and sensitive electrochemical biosensor for ATP based on the dual strategy integrating the cofactor-dependent enzymatic ligation reaction with self-cleaving DNAzyme-amplified electrochemical detection.

    PubMed

    Lu, Lu; Si, Jing Cao; Gao, Zhong Feng; Zhang, Yu; Lei, Jing Lei; Luo, Hong Qun; Li, Nian Bing

    2015-01-15

    A dual strategy that combines the adenosine triphosphate (ATP)-dependent enzymatic ligation reaction with self-cleaving DNAzyme-amplified electrochemical detection is employed to construct the biosensor. In this design, the methylene blue-labeled hairpin-structured DNA was self-assembled onto a gold electrode surface to prepare the modified electrode through the interaction of Au-S bond. In the procedure of ATP-dependent ligation reaction, when the specific cofactor ATP was added, the two split oligonucleotide fragments of 8-17 DNAzyme were linked by T4 DNA ligase and then released to hybridize with the labeled hairpin-structured DNA substrate. The linked 8-17 DNAzyme catalyzes the cleavage of the hairpin-structured substrate by the addition of Zn(2+), causing the methylene blue which contains high electrochemical activity to leave the surface of the gold electrode, therefore generating a dramatic decrease of electrochemical signal. The decrease of peak current was readily measured by square wave voltammetry and a relatively low detection limit (0.05 nM) was obtained with a linear response range from 0.1 to 1000 nM. By taking advantage of the highly specific cofactor dependence of the DNA ligation reaction, the proposed ligation-induced DNAzyme cascades demonstrate ultrahigh selectivity toward the target cofactor ATP. A catalytic and molecular beacons strategy is further adopted to amplify the electrochemical signal detection achieved by cycling and regenerating the 8-17 DNAzyme to realize enzymatic multiple turnover, thus one DNAzyme can catalyze the cleavage of several hairpin-structured substrates, which improves the sensitivity of the newly designed electrochemical sensing system. PMID:25048448

  15. Parallel Optical and Electrochemical DNA Detection

    NASA Astrophysics Data System (ADS)

    Knoll, Wolfgang; Liu, Jianyun; Niu, Lifang; Nielsen, Peter Eigil; Tiefenauer, Louis

    This contribution introduces strategies for the sensitive detection of oligonucleotides as bio-analytes binding from solution to a variety of probe architectures assembled at the (Au-) sensor surface. Detection principles based on surface plasmon optics and electrochemical techniques are compared. In particular, cyclic- and square wave voltammetry (SWV) are applied for the read-out of ferrocene redox labels conjugated to streptavidin that binds to the (biotinylated) DNA targets after hybridizing to the interfacial probe matrix of either DNA or peptide nucleic acid (PNA) strands. By employing streptavidin modified with fluorophores the identical sensor architecture can be used for the recording of hybridization reactions by surface plasmon fluorescence spectroscopy (SPFS). The Langmuir isotherms determined by both techniques, i.e., by SWV and SPFS, give virtually identical affinity constants KA, confirming that the mode of detection has no influence on the hybridization reaction. By using semiconducting nanoparticles as luminescence labels that can be tuned in their bandgap energies over a wide range of emission wavelengths surface plasmon fluorescence microscopy allows for the parallel read-out of multiple analyte binding events simultaneously.

  16. Self-interconnecting Pt nanowire network electrode for electrochemical amperometric biosensor.

    PubMed

    Wang, Shuqi; Xu, Li-Ping; Liang, Hai-Wei; Yu, Shu-Hong; Wen, Yongqiang; Wang, Shutao; Zhang, Xueji

    2015-07-14

    One-dimensional Pt nanostructures are of considerable interest for the development of highly stable and sensitive electrochemical sensors. This paper describes a self-interconnecting Pt nanowire network electrode (PtNNE) for the detection of hydrogen peroxide (H2O2) and glucose with ultrahigh sensitivity and stability. The as-prepared PtNNE consists of polycrystalline nanowires with high-index facets along the side surface which provides more active surface atoms on kinks and steps, those ultralong nanowires being interconnected with each other to form a free-standing network membrane. The excellent structural features of the PtNNE promoted its performance as a Pt-based electrochemical sensor both in terms of electrocatalytic activity and stability. Amperometric measurements towards hydrogen peroxide were performed; the PtNNE sensor showed an extremely high sensitivity of 1360 μA mM(-1) cm(-2). This excellent sensitivity is mainly attributed to the high-index facets of the nanowires resulting in their superior electrocatalytic activity towards H2O2, and the interconnected nanowire network forming an "electron freeway" transport model, which could provide multiple electron pathways and fast electron transport on the electrode, leading to rapid reaction and sensitive signal detection. The as-prepared PtNNE also holds promise as an oxidase-based biosensor. As a proof of concept, a PtNNE-based glucose biosensor also showed an outstanding sensitivity as high as 114 μA mM(-1) cm(-2), a low detection limit of 1.5 μM, and an impressive detection range from 5 μM to 30 mM. PMID:26083932

  17. Fluorescent magnetic bead-based mast cell biosensor for electrochemical detection of allergens in foodstuffs.

    PubMed

    Jiang, Donglei; Zhu, Pei; Jiang, Hui; Ji, Jian; Sun, Xiulan; Gu, Wenshu; Zhang, Genyi

    2015-08-15

    In this study, a novel electrochemical rat basophilic leukemia cell (RBL-2H3) cell sensor, based on fluorescent magnetic beads, has been developed for the detection and evaluation of different allergens in foodstuffs. Fluorescein isothiocyanate (FITC) was successfully fused inside the SiO2 layer of SiO2 shell-coated Fe3O4 nanoparticles, which was superior to the traditional Fe3O4@SiO2@FITC modification process. The as-synthesized fluorescent magnetic beads were then encapsulated with lipidosome to form cationic magnetic fluorescent nanoparticles (CMFNPs) for mast cell magnetofection. The CMFNPs were then characterized by SEM, TEM, VSM, FTIR, and XRD analyses, and transfected into RBL-2H3 cells through a highly efficient, lipid-mediated magnetofection procedure. Magnetic glassy carbon electrode (MGCE), which possesses excellent reproducibility and regeneration qualities, was then employed to adsorb the CMFNP-transfected RBL-2H3 cells activated by an allergen antigen for electrochemical assay. Results show that the exposure of model antigen-dinitrophenol-bovine serum albumin (DNP-BSA) to anti-DNP IgE-sensitized mast cells induced a robust and long-lasting electrochemical impedance signal in a dose-dependent manner. The detection limit was identified at 3.3×10(-4) ng/mL. To demonstrate the utility of this mast cell-based biosensor for detection of real allergens in foodstuffs, Anti-Pen a1 IgE and Anti-PV IgE-activated cells were employed to quantify both shrimp allergen tropomyosin (Pen a 1) and fish allergen parvalbumin (PV). Results show high detection accuracy for these targets, with a limit of 0.03 μg/mL (shrimp Pen a 1) and 0.16 ng/mL (fish PV), respectively. To this effect, we conclude the proposed method is a facile, highly sensitive, innovative electrochemical method for the evaluation of food allergens. PMID:25889258

  18. Development of a multilayered polymeric DNA biosensor using radio frequency technology with gold and magnetic nanoparticles.

    PubMed

    Yang, Cheng-Hao; Kuo, Long-Sheng; Chen, Ping-Hei; Yang, Chii-Rong; Tsai, Zuo-Min

    2012-01-15

    This study utilized the radio frequency (RF) technology to develop a multilayered polymeric DNA sensor with the help of gold and magnetic nanoparticles. The flexible polymeric materials, poly (p-xylylene) (Parylene) and polyethylene naphtholate (PEN), were used as substrates to replace the conventional rigid substrates such as glass and silicon wafers. The multilayered polymeric RF biosensor, including the two polymer layers and two copper transmission structure layers, was developed to reduce the total sensor size and further enhance the sensitivity of the biochip in the RF DNA detection. Thioglycolic acid (TGA) was used on the surface of the proposed biochip to form a thiolate-modified sensing surface for DNA hybridization. Gold nanoparticles (AuNPs) and magnetic nanoparticles (MNPs) were used to immobilize on the surface of the biosensor to enhance overall detection sensitivity. In addition to gold nanoparticles, the magnetic nanoparticles has been demonstrated the applicability for RF DNA detection. The performance of the proposed biosensor was evaluated by the shift of the center frequency of the RF biosensor because the electromagnetic characteristic of the biosensors can be altered by the immobilized multilayer nanoparticles on the biosensor. The experimental results show that the detection limit of the DNA concentration can reach as low as 10 pM, and the largest shift of the center frequency with triple-layer AuNPs and MNPs can approach 0.9 and 0.7 GHz, respectively. Such the achievement implies that the developed biosensor can offer an alternative inexpensive, disposable, and highly sensitive option for application in biomedicine diagnostic systems because the price and size of each biochip can be effectively reduced by using fully polymeric materials and multilayer-detecting structures. PMID:22093770

  19. Ultrasensitive electrochemical biosensing for DNA using quantum dots combined with restriction endonuclease.

    PubMed

    Zhang, Can; Lou, Jing; Tu, Wenwen; Bao, Jianchun; Dai, Zhihui

    2015-01-21

    A universal and sensitive electrochemical biosensing platform for the detection and identification of DNA using CdSe quantum dots (CdSe QDs) as signal markers was designed. The detection mechanism was based on the specific recognition of MspI endonuclease combined with the signal amplification of gold nanoparticles (AuNPs). MspI endonuclease could recognize its specific sequence in the double-strand DNA (dsDNA) and cleave the dsDNA fragments linked with CdSe QDs from the electrode. The remaining attached CdSe QDs can be easily read out by square-wave voltammetry using an electrodeposited bismuth (Bi) film-modified glass carbon electrode. The concentrations of target DNA could be simultaneously detected by the signal of metal markers. Using mycobacterium tuberculosis (Mtb) DNA as a model, under the optimal conditions, the proposed biosensor could detect Mtb DNA down to 8.7 × 10(-15) M with a linear range of 5 orders of magnitude (from 1.0 × 10(-14) to 1.0 × 10(-9) M) and discriminate mismatched DNA with high selectivity. This strategy presented a universal and convenient biosensing platform for DNA assay, and its satisfactory performances make it a potential candidate for the early diagnosis of gene-related diseases. PMID:25408952

  20. Use of 3,3',5,5' tetramethylbenzidine as new electrochemical indicator of DNA hybridization and its application in genossensor.

    PubMed

    Alves-Balvedi, R P; Caetano, L P; Madurro, J M; Brito-Madurro, A G

    2016-11-15

    Electrochemical tools are important biosensor platforms for disease diagnosis, due to their speediness, easiness, low cost and portability. However, for DNA detection, the use of indicators and/or intercalators is necessary to improve electrochemical sensitivity. Currently, ethidium bromide (EthBr) is the cheapest and most used DNA intercalators, but presents carcinogenic and teratogenic properties. Other indicators may be important for DNA photonic detection, and besides being more expensive, they behave similarly to EthBr. This investigation shows for the first time the use of tetramethylbenzidine(TMB) as a new remarkable non-carcinogenic DNA indicator for genosensing purposes, which may be used for nucleic acid detection of microorganisms, based on complementarity of base-pairing between probe and target molecules. The results indicate that TMB can be used as a new electrochemical indicator readily applicable in genosensors, which is able to detect the hybridization of single stranded DNA probe with its complementary target strand. An additional advantage of TMB, beside its non-genotoxicity, is the electrochemical reduction property, which prevents interference of serum components and other oxidative samples in the electrochemical analysis. PMID:27179563

  1. Selective detection of silver ions using mushroom-like polyaniline and gold nanoparticle nanocomposite-based electrochemical DNA sensor.

    PubMed

    Yang, Yanqin; Zhang, Shuai; Kang, Mengmeng; He, Linghao; Zhao, Jihong; Zhang, Hongzhong; Zhang, Zhihong

    2015-12-01

    A highly sensitive electrochemical DNA biosensor made of polyaniline (PANI) and gold nanoparticles (AuNPs) nanocomposite (AuNPs@PANI) has been used for the detection of trace concentration of Ag(+). In the presence of Ag(+), with the interaction of cytosine-Ag(+)-cytosine (C-Ag(+)-C), cytosine-rich DNA sequence immobilized onto the surface of AuNPs@PANI has a self-hybridization and then forms a duplex-like structure. The whole detection procedure of Ag(+) based on the developed biosensor was evaluated by electrochemical impedance spectroscopy. On semi-logarithmic plots of the log Ag(+) concentration versus peak current, the results show that the prepared biosensor can detect silver ions at a wide linear range of 0.01-100 nM (R = 0.9828) with a detection limit of 10 pM (signal/noise = 3). Moreover, the fabricated sensor exhibits good selectivity and repeatability. The detection of Ag(+) was determined by Ag(+) self-induced conformational change of DNA scaffold that involved only one oligonucleotide, showing its convenience and availability. PMID:26292168

  2. Detection of Neisseria meningitidis using surface plasmon resonance based DNA biosensor.

    PubMed

    Kaur, Gurpreet; Paliwal, Ayushi; Tomar, Monika; Gupta, Vinay

    2016-04-15

    Herein, we report the development of a surface plasmon resonance (SPR) based biosensor for the detection of Neisseria meningitidis DNA employing Kretschmann configuration. Highly c-axis oriented ZnO thin film of thickness 200nm was deposited on gold coated glass prisms by RF sputtering technique. Single stranded probe DNA was immobilized on the surface of ZnO thin film by physical adsorption method. SPR reflectance curves were recorded as a function of incident angle of He-Ne laser beam using a laboratory assembled SPR setup. The prepared biosensor exhibits a linear response towards target meningitidis DNA over the concentration range from 10 to 180 ng/μl with a high sensitivity of about 0.03°/(ng/μl) and a low limit of detection of 5 ng/μl. The SPR biosensor demonstrated high specificity and long shelf life thus, pointing towards a promising application in the field of meningitidis diagnosis. PMID:26599479

  3. Construction of titanium dioxide nanorod/graphite microfiber hybrid electrodes for a high performance electrochemical glucose biosensor.

    PubMed

    Zhang, Jian; Yu, Xin; Guo, Weibo; Qiu, Jichuan; Mou, Xiaoning; Li, Aixue; Liu, Hong

    2016-04-28

    The demand for a highly sensitive and selective glucose biosensor which can be used for implantable or on-time monitoring is constantly increasing. In this work, TiO2 nanorods were synthesized in situ on the surface of graphite microfibers to yield TiO2 nanorod/graphite microfiber hybrid electrodes. The TiO2 nanorods not only retain the high activity of the immobilized glucose molecule, but also promote the direct electron transfer process on the electrode surface. As a working electrode in an electrochemical glucose biosensor in a flowing system, the microfiber hybrid electrodes exhibit high sensitivity, selectivity and stability. Due to its simplicity, low cost, high stability, and unique morphology, the TiO2 nanorod/graphite microfiber hybrid electrode is expected to be an excellent candidate for an implantable biosensor or for in situ flow monitoring. PMID:27093292

  4. Recent progress in electrochemical biosensors based on phenylboronic acid and derivatives.

    PubMed

    Anzai, Jun-Ichi

    2016-10-01

    This review provides an overview of recent progress made in the development of electrochemical biosensors based on phenylboronic acid (PBA) and its derivatives. PBAs are known to selectively bind 1,2- and 1,3-diols to form negatively charged boronate esters in neutral aqueous media and have been used to construct electrochemical glucose sensors because of this selective binding. PBA-modified metal and carbon electrodes have been widely studied as voltammetric and potentiometric glucose sensors. In some cases, ferroceneboronic acid or ferrocene-modified phenylboronic acids are used as sugar-selective redox compounds. Another option for sensors using PBA-modified electrodes is potentiometric detection, in which the changes in surface potential of the electrodes are detected as an output signal. An ion-sensitive field effect transistor (FET) has been used as a signal transducer in potentiometric sensors. Glycoproteins, such as glycated hemoglobin (HbA1c), avidin, and serum albumin can also be detected by PBA-modified electrodes because they contain hydrocarbon chains on the surface. HbA1c sensors are promising alternatives to enzyme-based glucose sensors for monitoring blood glucose levels over the preceding 2-3months. In addition, PBA-modified electrodes can be used to detect a variety of compounds including hydroxy acids and fluoride (F(-)) ions. PBA-based F(-) ion sensors may be useful if reagentless sensors can be developed. PMID:27287174

  5. Fabrication and Electrochemical Characterization of Nanoporous Silicon Electrode for Amperometric Urea Biosensor

    NASA Astrophysics Data System (ADS)

    Yun, Donghwa; Song, Min-Jung; Hwang, Sungwoo; Hong, Suk-In

    2012-06-01

    We describe a new type of biosensor that employs a modified gold electrode based on nanoporous silicon (NPSi) for the electrochemical detection of urea. Urease (Urs) was covalently immobilized onto an Au/NPSi electrode functionalized with 3-mercaptopropionic acid (3-MPA). Amperometric calibration curves for both NPSi and planar silicon (PLSi)-based urea sensitive electrodes were compared in the range of 0.3 to 4.5 mM urea concentrations. The Michaelis-Menten constant (Km) was determined using the amperometric method. The electrochemical active area (Aea) of the 3-MPA/Au/NPSi electrode was evaluated using cyclic voltammetry (CV) and the result was compared with the 3-MPA/Au/PLSi electrode. Measured sensitivity of the Urs/SAMs/Au/NPSi electrode is ca. 2.05 µA mM-1 cm-2 and that of the Urs/SAMs/Au/PLSi electrode is ca. 1.10 µA mM-1 cm-2. About 1.8 times of sensitivity increase is obtained in the Au/NPSi electrode.

  6. Biocompatible enzymatic roller pens for direct writing of biocatalytic materials: "do-it-yourself" electrochemical biosensors.

    PubMed

    Bandodkar, Amay J; Jia, Wenzhao; Ramírez, Julian; Wang, Joseph

    2015-06-01

    The development of enzymatic-ink-based roller pens for direct drawing of biocatalytic sensors, in general, and for realizing renewable glucose sensor strips, in particular, is described. The resulting enzymatic-ink pen allows facile fabrication of high-quality inexpensive electrochemical biosensors of any design by the user on a wide variety of surfaces having complex textures with minimal user training. Unlike prefabricated sensors, this approach empowers the end user with the ability of "on-demand" and "on-site" designing and fabricating of biocatalytic sensors to suit their specific requirement. The resulting devices are thus referred to as "do-it-yourself" sensors. The bio-active pens produce highly reproducible biocatalytic traces with minimal edge roughness. The composition of the new enzymatic inks has been optimized for ensuring good biocatalytic activity, electrical conductivity, biocompati-bility, reproducible writing, and surface adherence. The resulting inks are characterized using spectroscopic, viscometric, electrochemical, thermal and microscopic techniques. Applicability to renewable blood glucose testing, epidermal glucose monitoring, and on-leaf phenol detection are demonstrated in connection to glucose oxidase and tyrosinase-based carbon inks. The "do-it-yourself" renewable glucose sensor strips offer a "fresh," reproducible, low-cost biocatalytic sensor surface for each blood test. The ability to directly draw biocatalytic conducting traces even on unconventional surfaces opens up new avenues in various sensing applications in low-resource settings and holds great promise for diverse healthcare, environmental, and defense domains. PMID:25721554

  7. Self-assembled magnetic bead chains for sensitivity enhancement of microfluidic electrochemical biosensor platforms.

    PubMed

    Armbrecht, L; Dincer, C; Kling, A; Horak, J; Kieninger, J; Urban, G

    2015-11-21

    In this paper, we present a novel approach to enhance the sensitivity of microfluidic biosensor platforms with self-assembled magnetic bead chains. An adjustable, more than 5-fold sensitivity enhancement is achieved by introducing a magnetic field gradient along a microfluidic channel by means of a soft-magnetic lattice with a 350 μm spacing. The alternating magnetic field induces the self-assembly of the magnetic beads in chains or clusters and thus improves the perfusion and active contact between the analyte and the beads. The soft-magnetic lattices can be applied independent of the channel geometry or chip material to any microfluidic biosensing platform. At the same time, the bead-based approach achieves chip reusability and shortened measurement times. The bead chain properties and the maximum flow velocity for bead retention were validated by optical microscopy in a glass capillary. The magnetic actuation system was successfully validated with a biotin-streptavidin model assay on a low-cost electrochemical microfluidic chip, fabricated by dry-film photoresist technology (DFR). Labelling with glucose oxidase (GOx) permits rapid electrochemical detection of enzymatically produced H2O2. PMID:26394820

  8. Integrated electrochemical biosensor based on algal metabolism for water toxicity analysis.

    PubMed

    Tsopela, A; Lale, A; Vanhove, E; Reynes, O; Séguy, I; Temple-Boyer, P; Juneau, P; Izquierdo, R; Launay, J

    2014-11-15

    An autonomous electrochemical biosensor with three electrodes integrated on the same silicon chip dedicated to the detection of herbicides in water was fabricated by means of silicon-based microfabrication technology. Platinum (Pt), platinum black (Pt Bl), tungsten/tungsten oxide (W/WO3) and iridium oxide (Pt/IrO2) working ultramicroelectrodes were developed. Ag/AgCl and Pt electrodes were used as reference and counter-integrated electrodes respectively. Physical vapor deposition (PVD) and electrodeposition were used for thin film deposition. The ultramicroelectrodes were employed for the detection of O2, H2O2 and pH related ions H3O(+)/OH(-), species taking part in photosynthetic and metabolic activities of algae. By measuring the variations in consumption-production rates of these electroactive species by algae, the quantity of herbicides present at trace level in the solution can be estimated. Fabricated ultramicroelectrodes were electrochemically characterized and calibrated. Pt Black ultramicroelectrodes exhibited the greatest sensitivity regarding O2 and H2O2 detection while Pt/IrO2 ultramicroelectrodes were more sensitive for pH measurement compared to W/WO3 ultramicroelectrodes for pH measurement. Bioassays were then conducted to detect traces of Diuron herbicide in water samples by evaluating disturbances in photosynthetic and metabolic activities of algae caused by this herbicide. PMID:24906088

  9. Zinc oxide inverse opal electrodes modified by glucose oxidase for electrochemical and photoelectrochemical biosensor.

    PubMed

    Xia, Lei; Song, Jian; Xu, Ru; Liu, Dali; Dong, Biao; Xu, Lin; Song, Hongwei

    2014-09-15

    The ZnO inverse opal photonic crystals (IOPCs) were synthesized by the sol-gel method using the polymethylmethacrylate (PMMA) as a template. For glucose detection, glucose oxidase (GOD) was further immobilized on the inwall and surface of the IOPCs. The biosensing properties toward glucose of the Nafion/GOD/ZnO IOPCs modified FTO electrodes were carefully studied and the results indicated that the sensitivity of ZnO IOPCs modified electrode was 18 times than reference electrode due to the large surface area and uniform porous structure of ZnO IOPCs. Moreover, photoelectrochemical detection for glucose using the electrode was realized and the sensitivity approached to 52.4 µA mM(-1) cm(-2), which was about four times to electrochemical detection (14.1 µA mM(-1) cm(-2)). It indicated that photoelectrochemical detection can highly improve the sensor performance than conventional electrochemical method. It also exhibited an excellent anti-interference property and a good stability at the same time. This work provides a promising approach for realizing excellent photoelectrochemical biosensor of similar semiconductor photoelectric material. PMID:24752145

  10. Construction of titanium dioxide nanorod/graphite microfiber hybrid electrodes for a high performance electrochemical glucose biosensor

    NASA Astrophysics Data System (ADS)

    Zhang, Jian; Yu, Xin; Guo, Weibo; Qiu, Jichuan; Mou, Xiaoning; Li, Aixue; Liu, Hong

    2016-04-01

    The demand for a highly sensitive and selective glucose biosensor which can be used for implantable or on-time monitoring is constantly increasing. In this work, TiO2 nanorods were synthesized in situ on the surface of graphite microfibers to yield TiO2 nanorod/graphite microfiber hybrid electrodes. The TiO2 nanorods not only retain the high activity of the immobilized glucose molecule, but also promote the direct electron transfer process on the electrode surface. As a working electrode in an electrochemical glucose biosensor in a flowing system, the microfiber hybrid electrodes exhibit high sensitivity, selectivity and stability. Due to its simplicity, low cost, high stability, and unique morphology, the TiO2 nanorod/graphite microfiber hybrid electrode is expected to be an excellent candidate for an implantable biosensor or for in situ flow monitoring.The demand for a highly sensitive and selective glucose biosensor which can be used for implantable or on-time monitoring is constantly increasing. In this work, TiO2 nanorods were synthesized in situ on the surface of graphite microfibers to yield TiO2 nanorod/graphite microfiber hybrid electrodes. The TiO2 nanorods not only retain the high activity of the immobilized glucose molecule, but also promote the direct electron transfer process on the electrode surface. As a working electrode in an electrochemical glucose biosensor in a flowing system, the microfiber hybrid electrodes exhibit high sensitivity, selectivity and stability. Due to its simplicity, low cost, high stability, and unique morphology, the TiO2 nanorod/graphite microfiber hybrid electrode is expected to be an excellent candidate for an implantable biosensor or for in situ flow monitoring. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr01360k

  11. Fabrication and functionalization of PCB gold electrodes suitable for DNA-based electrochemical sensing.

    PubMed

    Salvo, P; Henry, O Y F; Dhaenens, K; Acero Sanchez, J L; Gielen, A; Werne Solnestam, B; Lundeberg, J; O'Sullivan, C K; Vanfleteren, J

    2014-01-01

    The request of high specificity and selectivity sensors suitable for mass production is a constant demand in medical research. For applications in point-of-care diagnostics and therapy, there is a high demand for low cost and rapid sensing platforms. This paper describes the fabrication and functionalization of gold electrodes arrays for the detection of deoxyribonucleic acid (DNA) in printed circuit board (PCB) technology. The process can be implemented to produce efficiently a large number of biosensors. We report an electrolytic plating procedure to fabricate low-density gold microarrays on PCB suitable for electrochemical DNA detection in research fields such as cancer diagnostics or pharmacogenetics, where biosensors are usually targeted to detect a small number of genes. PCB technology allows producing high precision, fast and low cost microelectrodes. The surface of the microarray is functionalized with self-assembled monolayers of mercaptoundodecanoic acid or thiolated DNA. The PCB microarray is tested by cyclic voltammetry in presence of 5 mM of the redox probe K3Fe(CN6) in 0.1 M KCl. The voltammograms prove the correct immobilization of both the alkanethiol systems. The sensor is tested for detecting relevant markers for breast cancer. Results for 5 nM of the target TACSTD1 against the complementary TACSTD1 and non-complementary GRP, MYC, SCGB2A1, SCGB2A2, TOP2A probes show a remarkable detection limit of 0.05 nM and a high specificity. PMID:24948454

  12. Sensitivity Enhancement of Bead-based Electrochemical Impedance Spectroscopy (BEIS) biosensor by electric field-focusing in microwells.

    PubMed

    Shin, Kyeong-Sik; Ji, Jae Hoon; Hwang, Kyo Seon; Jun, Seong Chan; Kang, Ji Yoon

    2016-11-15

    This paper reports a novel electrochemical impedance spectroscopy (EIS) biosensors that uses magnetic beads trapped in a microwell array to improve the sensitivity of conventional bead-based EIS (BEIS) biosensors. Unloading the previously measured beads by removing the magnetic bar enables the BEIS sensor to be used repeatedly by reloading it with new beads. Despite its recyclability, the sensitivity of conventional BEIS biosensors is so low that it has not attracted much attentions from the biosensor industry. We significantly improved the sensitivity of the BEIS system by introducing of a microwell array that contains two electrodes (a working electrode and a counter electrode) to concentrate the electric field on the surfaces of the beads. We confirmed that the performance of the BEIS sensor in a microwell array using an immunoassay of prostate specific antigen (PSA) in PBS buffer and human plasma. The experimental results showed that a low concentration of PSA (a few tens or hundreds of fg/mL) were detectable as a ratio of the changes in the impedance of the PBS buffer or in human plasma. Therefore, our BEIS sensor with a microwell array could be a promising platform for low cost, high-performance biosensors for applications that require high sensitivity and recyclability. PMID:27152445

  13. DNA probe functionalized QCM biosensor based on gold nanoparticle amplification for Bacillus anthracis detection.

    PubMed

    Hao, Rong-Zhang; Song, Hong-Bin; Zuo, Guo-Min; Yang, Rui-Fu; Wei, Hong-Ping; Wang, Dian-Bing; Cui, Zong-Qiang; Zhang, ZhiPing; Cheng, Zhen-Xing; Zhang, Xian-En

    2011-04-15

    The rapid detection of Bacillus anthracis, the causative agent of anthrax disease, has gained much attention since the anthrax spore bioterrorism attacks in the United States in 2001. In this work, a DNA probe functionalized quartz crystal microbalance (QCM) biosensor was developed to detect B. anthracis based on the recognition of its specific DNA sequences, i.e., the 168 bp fragment of the Ba813 gene in chromosomes and the 340 bp fragment of the pag gene in plasmid pXO1. A thiol DNA probe was immobilized onto the QCM gold surface through self-assembly via Au-S bond formation to hybridize with the target ss-DNA sequence obtained by asymmetric PCR. Hybridization between the target DNA and the DNA probe resulted in an increase in mass and a decrease in the resonance frequency of the QCM biosensor. Moreover, to amplify the signal, a thiol-DNA fragment complementary to the other end of the target DNA was functionalized with gold nanoparticles. The results indicate that the DNA probe functionalized QCM biosensor could specifically recognize the target DNA fragment of B. anthracis from that of its closest species, such as Bacillus thuringiensis, and that the limit of detection (LOD) reached 3.5 × 10(2)CFU/ml of B. anthracis vegetative cells just after asymmetric PCR amplification, but without culture enrichment. The DNA probe functionalized QCM biosensor demonstrated stable, pollution-free, real-time sensing, and could find application in the rapid detection of B. anthracis. PMID:21315574

  14. DNA-based biosensor for comparative study of catalytic effect of transition metals on autoxidation of sulfite.

    PubMed

    Ensafi, Ali A; Heydari-Bafrooei, Esmaeil; Rezaei, Behzad

    2013-01-15

    The transition metal-catalyzed oxidation of sulfur(IV) oxides has been known for more than 100 years. However, to the best of the authors' knowledge, no electrochemical quantitative study has yet been carried out to determine its nature. In view of the transition metal catalyzed oxidation of sulfur(IV) oxides, a series of radicals are involved in the overall reaction process whereby the sulfite, in the presence of transition metals, may cause damages to DNA through the generation of these highly reactive species. In the present work, {MWCNTs-PDDA/DNA}(2) layer-by-layer (LBL) films were prepared to detect DNA damage induced by radicals generated from sulfite autoxidation using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The change in the peak potential separation (ΔE(p)) and charge transfer resistance (R(p)) after incubation of the DNA biosensor in the damaging solution for a certain time was used as indicators of DNA damage. It was found that sulfite in the presence of Co(II), Cu(II), Cr(VI), Fe(III), and Mn(II) caused damage to DNA while neither sulfite alone nor metal ions alone did have the same effect. The results suggest that sulfite is rapidly autoxidized in the presence of Co(II), Cu(II), Cr(VI), Fe(III), and Mn(II), producing radicals that cause the DNA damage. These radicals can be ranked in a descending order of their ability to induce DNA damage with sulfite as follows: Fe(III) > Co(II) > Cu(II) > Cr(VI) > Mn(II). The DNA damage induced by sulfite plus Co(II), Cr(VI), and Fe(III) was inhibited by primary alcohols, but they were not when superoxide dismutase (SOD) and tert-butyl alcohol were used. Comparison of methods used to determine the minimum concentration of a transition metal for sulfite induced DNA damage revealed that electrochemical impedance spectroscopy and cyclic voltammetry outperformed the quantitative comparison of different reagents. PMID:23244055

  15. A novel self-powered and sensitive label-free DNA biosensor in microbial fuel cell.

    PubMed

    Asghary, Maryam; Raoof, Jahan Bakhsh; Rahimnejad, Mostafa; Ojani, Reza

    2016-08-15

    In this work, a novel self-powered, sensitive, low-cost, and label-free DNA biosensor is reported by applying a two-chambered microbial fuel cell (MFC) as a power supply. A graphite electrode and an Au nanoparticles modified graphite electrode (AuNP/graphite electrode) were used as anode and cathode in the MFC system, respectively. The active biocatalyst in the anodic chamber was a mixed culture of microorganisms. The sensing element of the biosensor was fabricated by the well-known Au-thiol binding the ssDNA probe on the surface of an AuNP/graphite cathode. Electrons produced by microorganisms were transported from the anode to the cathode through an external circuit, which could be detected by the terminal multi-meter detector. The difference between power densities of the ssDNA probe modified cathode in the absence and presence of complementary sequence served as the detection signal of the DNA hybridization with detection limit of 3.1nM. Thereafter, this biosensor was employed for diagnosis and determination of complementary sequence in a human serum sample. The hybridization specificity studies further revealed that the developed DNA biosensor could distinguish fully complementary sequences from one-base mismatched and non-complementary sequences. PMID:27085948

  16. Detection of DNA and Protein using CVD Graphene-channel FET Biosensors

    NASA Astrophysics Data System (ADS)

    Sebastian, Abhilash; Kakatkar, Aniket; de Alba, Roberto; Craighead, Harold; Parpia, Jeevak

    2015-03-01

    Graphene channel field-effect biosensors are demonstrated for detecting the binding of double-stranded DNA and poly-l-lysine. Sensors consist of CVD graphene transferred using a clean, etchant-free transfer method. The presence of DNA and poly-l-lysine are detected by the change in the Dirac Voltage (the voltage at which the graphene's resistance peaks) of the graphene transistor. Sensors show large shifts in the Dirac voltage ~ 17 V after exposure to ~ 580 pM of poly-l-lysine and ~ 14 V upon exposure to 300 pM of DNA. The polarity of the response changes to positive direction with poly-l-lysine and negative direction with DNA. Sensors show detection limits of 8 pM for 48.5 kbp DNA and 11 pM for poly-l-lysine. The biosensors are easy to fabricate, reusable and are promising as sensors of a wide variety of charged biomolecules.

  17. RCA-Based Biosensor for Electrical and Colorimetric Detection of Pathogen DNA

    NASA Astrophysics Data System (ADS)

    Jeong, Jaepil; Kim, Hyejin; Lee, Dong Jun; Jung, Byung Jun; Lee, Jong Bum

    2016-05-01

    For the diagnosis and prevention of diseases, a range of strategies for the detection of pathogens have been developed. In this study, we synthesized the rolling circle amplification (RCA)-based biosensor that enables detection of pathogen DNA in two analytical modes. Only in the presence of the target DNA, the template DNA can be continuously polymerized by simply carrying out RCA, which gives rise to a change of surface structure of Au electrodes and the gap between the electrodes. Electrical signal was generated after introducing hydrogen tetrachloroaurate (HAuCl4) to the DNA-coated biosensor for the improvement of the conductivity of DNA, which indicates that the presence of the pathogen DNA can be detected in an electrical approach. Furthermore, the existence of the target DNA was readily detected by the naked eyes through change in colors of the electrodes from bright yellow to orange-red after RCA reaction. The RCA-based biosensor offers a new platform for monitoring of pathogenic DNA with two different detection modes in one system.

  18. RCA-Based Biosensor for Electrical and Colorimetric Detection of Pathogen DNA.

    PubMed

    Jeong, Jaepil; Kim, Hyejin; Lee, Dong Jun; Jung, Byung Jun; Lee, Jong Bum

    2016-12-01

    For the diagnosis and prevention of diseases, a range of strategies for the detection of pathogens have been developed. In this study, we synthesized the rolling circle amplification (RCA)-based biosensor that enables detection of pathogen DNA in two analytical modes. Only in the presence of the target DNA, the template DNA can be continuously polymerized by simply carrying out RCA, which gives rise to a change of surface structure of Au electrodes and the gap between the electrodes. Electrical signal was generated after introducing hydrogen tetrachloroaurate (HAuCl4) to the DNA-coated biosensor for the improvement of the conductivity of DNA, which indicates that the presence of the pathogen DNA can be detected in an electrical approach. Furthermore, the existence of the target DNA was readily detected by the naked eyes through change in colors of the electrodes from bright yellow to orange-red after RCA reaction. The RCA-based biosensor offers a new platform for monitoring of pathogenic DNA with two different detection modes in one system. PMID:27142880

  19. An electrochemical microRNAs biosensor with the signal amplification of alkaline phosphatase and electrochemical-chemical-chemical redox cycling.

    PubMed

    Xia, Ning; Zhang, Youjuan; Wei, Xin; Huang, Yaping; Liu, Lin

    2015-06-01

    MicroRNAs (MiRNAs) have been regarded as clinically important biomarkers and drug discovery targets. In this work, we reported a simple and ultrasensitive electrochemical method for miRNAs detection based on single enzyme amplification and electrochemical-chemical-chemical (ECC) redox cycling. Specifically, upon contact with the target miRNAs, the hairpin structure of biotinylated DNA immobilized on gold electrode was destroyed and the biotin group in DNA was forced away from the electrode surface, allowing for the coupling of streptavidin-conjugated alkaline phosphatase (SA-ALP). Then, ascorbic acid (AA, the enzymatic product of ALP) triggered the ECC redox cycling with ferrocene methanol (FcM) and tris(2-carboxyethyl)phosphine (TCEP) as the redox mediator and the chemical reducing reagent, respectively. The method was more sensitive than that with horseradish peroxidase (HRP) or glucose oxidase (GOx) triggered recycling since one ALP molecule captured by one target miRNA molecule promoted the production of thousands of AA. Analytical merits (e.g., detection limit, dynamic range, specificity, regeneration and reproducibility) were evaluated. The feasibility of the method for analysis of miRNA-21 in human serum has also been demonstrated. PMID:26002330

  20. Electrochemical probe for the monitoring of DNA-protein interactions.

    PubMed

    Meunier-Prest, Rita; Bouyon, Alice; Rampazzi, Eve; Raveau, Suzanne; Andreoletti, Pierre; Cherkaoui-Malki, Mustapha

    2010-08-15

    Self-assembly of thiol-terminated oligonucleotides on gold substrates provides a convenient way for DNA-functionalized surfaces. Here we describe the development of an electrochemical assay for the detection of DNA-protein interactions based on the modification of the electrochemical response of methylene blue (MB) intercalated in the DNA strands. Using a functionalized electrode with double stranded DNA carrying T3 RNA polymerase binding sequence, we show a substantial attenuation of the current upon the DNA-protein interaction. Moreover, a Langmuir binding isotherm for T3 RNA polymerase (T3 Pol) gives a dissociation constant K(D) equal to 0.46+/-0.23 microM. Such value is 100 times lower than the calculated K(D) for the non-specific interaction of bovine serum albumin (BSA) with T3 Pol promoter. In addition, the use of the T7 RNA polymerase (T7 Pol) promoter instead of the T3 Pol promoter induces an increase of K(D) from 0.46 microM to more than 25 microM. Accordingly, this strong decrease in the affinity of T3 Pol towards an off-target DNA promoter reveals an electrochemical sequence-specific discrimination of DNA-protein interactions. In conclusion, our results show that the developed electrochemical test allows the monitoring of DNA-protein interactions with high specificity and with an in situ protein detection threshold at a nanomolar range. PMID:20447818

  1. A label-free ultrasensitive electrochemical DNA sensor based on thin-layer MoS2 nanosheets with high electrochemical activity.

    PubMed

    Wang, Xinxing; Nan, Fuxin; Zhao, Jinlong; Yang, Tao; Ge, Tong; Jiao, Kui

    2015-02-15

    A label-free and ultrasensitive electrochemical DNA biosensor, based on thin-layer molybdenum disulfide (MoS2) nanosheets sensing platform and differential pulse voltammetry detection, is constructed in this paper. The thin-layer MoS2 nanosheets were prepared via a simple ultrasound exfoliation method from bulk MoS2, which is simpler and no distortion compared with mechanical cleavage and lithium intercalation. Most importantly, this procedure allows the formation of MoS2 with enhanced electrochemical activity. Based on the high electrochemical activity and different affinity toward ssDNA versus dsDNA of the thin-layer MoS2 nanosheets sensing platform, the tlh gene sequence assay can be performed label-freely from 1.0 × 10(-16)M to 1.0 × 10(-10)M with a detection limit of 1.9 × 10(-17)M. Without labeling and the use of amplifiers, the detection method described here not only expands the application of MoS2, but also offers a viable alternative for DNA analysis, which has the priority in sensitivity, simplicity, and costs. Moreover, the proposed sensing platform has good electrocatalytic activity, and can be extended to detect more targets, such as guanine and adenine, which further expands the application of MoS2. PMID:25262063

  2. Novel electrochemical sensor based on functionalized graphene for simultaneous determination of adenine and guanine in DNA.

    PubMed

    Huang, Ke-Jing; Niu, De-Jun; Sun, Jun-Yong; Han, Cong-Hui; Wu, Zhi-Wei; Li, Yan-Li; Xiong, Xiao-Qin

    2011-02-01

    A nano-material carboxylic acid functionalized graphene (graphene-COOH) was prepared and used to construct a novel biosensor for the simultaneous detection of adenine and guanine. The direct electrooxidation behaviors of adenine and guanine on the graphene-COOH modified glassy carbon electrode (graphene-COOH/GCE) were carefully investigated by cyclic voltammetry and differential pulse voltammetry. The results indicated that both adenine and guanine showed the increase of the oxidation peak currents with the negative shift of the oxidation peak potentials in contrast to that on the bare glassy carbon electrode. The electrochemical parameters of adenine and guanine on the graphene-COOH/GCE were calculated and a simple and reliable electroanalytical method was developed for the detection of adenine and guanine, respectively. The modified electrode exhibited good behaviors in the simultaneous detection of adenine and guanine with the peak separation as 0.334V. The detection limit for individual determination of guanine and adenine was 5.0×10(-8)M and 2.5×10(-8)M (S/N=3), respectively. Furthermore, the measurements of thermally denatured single-stranded DNA were carried out and the value of (G+C)/(A+T) of single-stranded DNA was calculated as 0.80. The biosensor exhibited some advantages, such as simplicity, rapidity, high sensitivity, good reproducibility and long-term stability. PMID:21050729

  3. Study of interactions between DNA and aflatoxin B1 using electrochemical and fluorescence methods.

    PubMed

    Banitaba, Mohammad Hossein; Davarani, Saied Saeed Hosseiny; Mehdinia, Ali

    2011-04-15

    In this study, a carbon paste electrode modified with N-butylpyridinium hexafluorophosphate (BPPF(6)) ionic liquid and DNA was introduced as an electrochemical biosensor to study the interaction between DNA and aflatoxin B1 molecules. For this purpose, variations in oxidation peak current of guanine in various concentrations of aflatoxin B1 were measured by using the differential pulse voltammetry (DPV) method. According to this study, the binding constant of DNA-aflatoxin B1 was found to be 3.5×10(6)M(-1). This modified electrode was also used for determination of low concentrations of aflatoxin B1 by using differential pulse voltammetry. A linear dynamic range from 8.00×10(-8) to 5.91×10(-7)M and a limit of detection of 2.00×10(-8)M resulted from DPV measurements. To confirm our results, a fluorescence study was also performed. It resulted in a binding constant of 2.8×10(6)M(-1), which is in good agreement with that obtained from electrochemical study. PMID:21238426

  4. Investigation of Hemoglobin/Gold Nanoparticle Heterolayer on Micro-Gap for Electrochemical Biosensor Application

    PubMed Central

    Lee, Taek; Kim, Tae-Hyung; Yoon, Jinho; Chung, Yong-Ho; Lee, Ji Young; Choi, Jeong-Woo

    2016-01-01

    In the present study, we fabricated a hemoglobin/gold nanoparticle (Hb/GNP) heterolayer immobilized on the Au micro-gap to confirm H2O2 detection with a signal-enhancement effect. The hemoglobin which contained the heme group catalyzed the reduction of H2O2. To facilitate the electron transfer between hemoglobin and Au micro-gap electrode, a gold nanoparticle was introduced. The Au micro-gap electrode that has gap size of 5 µm was fabricated by conventional photolithographic technique to locate working and counter electrodes oppositely in a single chip for the signal sensitivity and reliability. The hemoglobin was self-assembled onto the Au surface via chemical linker 6-mercaptohexanoic acid (6-MHA). Then, the gold nanoparticles were adsorbed onto hemoglobin/6-MHA heterolayers by the layer-by-layer (LbL) method. The fabrication of the Hb/GNP heterolayer was confirmed by atomic force microscopy (AFM) and surface-enhanced Raman spectroscopy (SERS). The redox property and H2O2 detection of Hb/GNP on the micro-gap electrode was investigated by a cyclic voltammetry (CV) experiment. Taken together, the present results show that the electrochemical signal-enhancement effect of a hemoglobin/nanoparticle heterolayer was well confirmed on the micro-scale electrode for biosensor applications. PMID:27171089

  5. A Low-Cost Smartphone-Based Electrochemical Biosensor for Point-of-Care Diagnostics

    PubMed Central

    Sun, Alexander; Wambach, Travis; Venkatesh, A. G.; Hall, Drew A.

    2015-01-01

    This paper describes the development of a smartphone-based electrochemical biosensor module. The module contains a low power potentiostat that interfaces and harvests power from a smartphone through the phone’s audio jack. A prototype with two different potentiostat designs was constructed and used to conduct proof of concept cyclic voltammetry experiments with potassium ferro-/ferricyanide (K4[Fe(CN)6] / K3[Fe(CN)6]) in a side-by-side comparison with a laboratory grade instrument. Results show that the module functions within the available power budget and that the recovered voltammogram data matches well with the data from an expensive bench top tool. Excluding the loses from supply rectification and regulation, the module consumes either 5.7 mW or 4.3 mW peak power, depending on which of the two discussed potentiostat designs is used. At single quantity pricing, the hardware for the prototype device costs less than $30. PMID:26097899

  6. An immobilization-free electrochemical impedance biosensor based on duplex-specific nuclease assisted target recycling for amplified detection of microRNA.

    PubMed

    Zhang, Jing; Wu, Dong-Zhi; Cai, Shu-Xian; Chen, Mei; Xia, Yao-Kun; Wu, Fang; Chen, Jing-Hua

    2016-01-15

    An immobilization-free electrochemical impedance biosensor for microRNA detection was developed in this work, which was based on both the duplex-specific nuclease assisted target recycling (DSNATR) and capture probes (Cps) enriched from the solution to electrode surface via magnetic beads (MBs). In the absence of miR-21, Cps cannot be hydrolyzed due to the low activity of duplex-specific nuclease (DSN) against ssDNA. Therefore, the intact Cps could be attached to the surface of magnetic glass carbon electrode (MGCE), resulting in a compact negatively charged layer as well as a large charge-transfer resistance. While in the presence of miR-21, it hybridized with Cp to form a DNA-RNA heteroduplex. Due to the considerable cleavage preference for DNA in DNA-RNA hybrids, DSN hydrolyzed the target-binding part of the Cp while liberating the intact miR-21 to hybridize with a new Cp and initiate the second cycle of hydrolysis. In this way, a single miR-21 was able to trigger the permanent hydrolysis of multiple Cps. Finally, all Cps were digested. Thus, the negatively charged layer could not be formed, resulting in a small charge-transfer resistance. By employing the above strategy, the proposed biosensor achieved ultrahigh sensitivity toward miR-21 with a detection limit of 60aM. Meanwhile, the method showed little cross-hybridization among the closely related miRNA family members even at the single-base-mismatched level. Successful attempts were made in applying the approach to detect miR-21 in human serum samples of breast cancer patients. PMID:26363493

  7. Co-immobilization of glucoamylase and glucose oxidase for electrochemical sequential enzyme electrode for starch biosensor and biofuel cell.

    PubMed

    Lang, Qiaolin; Yin, Long; Shi, Jianguo; Li, Liang; Xia, Lin; Liu, Aihua

    2014-01-15

    A novel electrochemical sequential biosensor was constructed by co-immobilizing glucoamylase (GA) and glucose oxidase (GOD) on the multi-walled carbon nanotubes (MWNTs)-modified glassy carbon electrode (GCE) by chemical crosslinking method, where glutaraldehyde and bovine serum albumin was used as crosslinking and blocking agent, respectively. The proposed biosensor (GA/GOD/MWNTs/GCE) is capable of determining starch without using extra sensors such as Clark-type oxygen sensor or H2O2 sensor. The current linearly decreased with the increasing concentration of starch ranging from 0.005% to 0.7% (w/w) with the limit of detection of 0.003% (w/w) starch. The as-fabricated sequential biosensor can be applicable to the detection of the content of starch in real samples, which are in good accordance with traditional Fehling's titration. Finally, a stable starch/O2 biofuel cell was assembled using the GA/GOD/MWNTs/GCE as bioanode and laccase/MWNTs/GCE as biocathode, which exhibited open circuit voltage of ca. 0.53 V and the maximum power density of 8.15 μW cm(-2) at 0.31 V, comparable with the other glucose/O2 based biofuel cells reported recently. Therefore, the proposed biosensor exhibited attractive features such as good stability in weak acidic buffer, good operational stability, wide linear range and capable of determination of starch in real samples as well as optimal bioanode for the biofuel cell. PMID:23954673

  8. Thin-film microelectric arrays for amperometric enzyme biosensors with electrochemically synthesized glucose oxidase-polyaniline membrane

    NASA Astrophysics Data System (ADS)

    Dzyadevich, Sergei V.; Rossokhaty, Victor K.; Shram, Nataly; Shul'ga, Alexander A.; Soldatkin, Alexey P.; Strikha, Vitaly I.

    1994-10-01

    An amperometric glucose biosensor was fabricated by the electrochemical polymerization of aniline onto a gold electrodes in presence of glucose oxidase in phosphate buffer solution, pH 7.0. Aniline is easily polymerized forming a thin film, which adheres tightly on the electrodes surface. During the electropolymerization process the enzyme was entrapped into the polyaniline film being able to catalyze the hydrolysis of glucose. The experiments were performed to determine the optimal condition for polyaniline-glucose oxidase film preparation. Glucose can be determined by the biosensor in the concentration range 10-4 M to 2 X 10-2 M. The linearity of the biosensor response was observed from 2 X 10-4 M to 6 X 10-3 M glucose, which demonstrated that the internal diffusion of substrates and products of reaction through the polyaniline layer to the electrodes surface was the main limiting factor controlling the response value. The method of electropolymerization was found to have several advantage in comparison with other approaches especially for further mass manufacturing of the biosensors.

  9. Effect of platinum nanoparticle deposition parameters on hydrogen peroxide transduction for applications in wearable electrochemical glucose biosensors

    NASA Astrophysics Data System (ADS)

    Cargill, Allison A.; Neil, Kathrine M.; Hondred, John A.; McLamore, Eric S.; Claussen, Jonathan C.

    2016-05-01

    Enhanced interest in wearable biosensor technology over the past decade is directly related to the increasing prevalence of diabetes and the associated requirement of daily blood glucose monitoring. In this work we investigate the platinum-carbon transduction element used in traditional first-generation glucose biosensors which rely on the concentration of hydrogen peroxide produced by the glucose-glucose oxidase binding scheme. We electrodeposit platinum nanoparticles on a commercially-available screen printed carbon electrode by stepping an applied current between 0 and 7.12 mA/cm2 for a varying number of cycles. Next, we examine the trends in deposition and the effect that the number of deposition cycles has on the sensitivity of electrochemical glucose sensing. Results from this work indicate that applying platinum nanoparticles to screen printed carbon via electrodeposition from a metal salt solution improves overall biosensor sensitivity. This work also pinpoints the amount of platinum (i.e., number of deposition cycles) that maximizes biosensor sensitivity in an effort to minimize the use of the precious metals, viz., platinum, in electrode fabrication. In summary, this work quantifies the relationship between platinum electrodeposition and sensor performance, which is crucial in designing and producing cost-effective sensors.

  10. Electrochemical lactate biosensor based upon chitosan/carbon nanotubes modified screen-printed graphite electrodes for the determination of lactate in embryonic cell cultures.

    PubMed

    Hernández-Ibáñez, Naiara; García-Cruz, Leticia; Montiel, Vicente; Foster, Christopher W; Banks, Craig E; Iniesta, Jesús

    2016-03-15

    l-lactate is an essential metabolite present in embryonic cell culture. Changes of this important metabolite during the growth of human embryo reflect the quality and viability of the embryo. In this study, we report a sensitive, stable, and easily manufactured electrochemical biosensor for the detection of lactate within embryonic cell cultures media. Screen-printed disposable electrodes are used as electrochemical sensing platforms for the miniaturization of the lactate biosensor. Chitosan/multi walled carbon nanotubes composite have been employed for the enzymatic immobilization of the lactate oxidase enzyme. This novel electrochemical lactate biosensor analytical efficacy is explored towards the sensing of lactate in model (buffer) solutions and is found to exhibit a linear response towards lactate over the concentration range of 30.4 and 243.9 µM in phosphate buffer solution, with a corresponding limit of detection (based on 3-sigma) of 22.6 µM and exhibits a sensitivity of 3417 ± 131 µAM(-1) according to the reproducibility study. These novel electrochemical lactate biosensors exhibit a high reproducibility, with a relative standard deviation of less than 3.8% and an enzymatic response over 82% after 5 months stored at 4 °C. Furthermore, high performance liquid chromatography technique has been utilized to independently validate the electrochemical lactate biosensor for the determination of lactate in a commercial embryonic cell culture medium providing excellent agreement between the two analytical protocols. PMID:26579934

  11. Determination of Parathion and Carbaryl Pesticides in Water and Food Samples Using a Self Assembled Monolayer/Acetylcholinesterase Electrochemical Biosensor

    PubMed Central

    Pedrosa, Valber A.; Caetano, Josiane; Machado, Sergio A. S.; Bertotti, Mauro

    2008-01-01

    An acetylcholinesterase (AchE) based amperometric biosensor was developed by immobilisation of the enzyme onto a self assembled modified gold electrode. Cyclic voltammetric experiments performed with the SAM-AchE biosensor in phosphate buffer solutions (pH = 7.2) containing acetylthiocholine confirmed the formation of thiocholine and its electrochemical oxidation at Ep = 0.28 V vs Ag/AgCl. An indirect methodology involving the inhibition effect of parathion and carbaryl on the enzymatic reaction was developed and employed to measure both pesticides in spiked natural water and food samples without pre-treatment or pre-concentration steps. Values higher than 91-98.0% in recovery experiments indicated the feasibility of the proposed electroanalytical methodology to quantify both pesticides in water or food samples. HPLC measurements were also performed for comparison and confirmed the values measured amperometrically.

  12. SiO2 nanoparticles modified CPE as a biosensor for determination of i-motif DNA/Tamoxifen interaction.

    PubMed

    Heydari, Elham; Raoof, Jahan Bakhsh; Ojani, Reza; Bagheryan, Zahra

    2016-08-01

    Cytosine-rich DNA sequences can form a highly ordered structure known as i-motif in slightly acidic solutions. The stability of the folded i-motif structure is a good strategy to inhibit the telomerase reaction in cancer cells. The electrochemical biosensor was prepared by modifying carbon paste electrode with SiO2 nanoparticles to investigate drugs which can stabilize this structure. Tamoxifen (Tam), an antiestrogen hormonal agent for treatment of breast cancer, was chosen as the model ligand and its interaction with i-motif structure was examined. The interaction between i-motif DNA and Tam was studied in PBS buffer and [Fe(CN)6](3-) through the cyclic voltammetry and square wave voltammetry methods. The oxidation peak of Tam, due to the i-motif DNA/Tam interaction, was observed after i-motif immobilized on the surface of the electrode. The i-motif formation was investigated by circular dichroism spectroscopy and the results showed that this structure can certainly be made with pH around 4.5, but its stability reduced by going to the more alkaline pH. The selectivity which was studied in the presence of complementary strand demonstrated that i-motif structure could be stabilized in acidic pH even in the presence of its complementary strand. PMID:27151665

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

    PubMed Central

    Wang, Qingxiang; Gao, Feng; Ni, Jiancong; Liao, Xiaolei; Zhang, Xuan; Lin, Zhenyu

    2016-01-01

    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. PMID:26931160

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

    PubMed

    Wang, Qingxiang; Gao, Feng; Ni, Jiancong; Liao, Xiaolei; Zhang, Xuan; Lin, Zhenyu

    2016-01-01

    An ultrasensitive DNA biosensor has been developed through in-situ labeling of electroactive melamine-Cu(2+) complex (Mel-Cu(2+)) 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 Cu(2+) were assembled on the AuNPs surface through Au-N bond and Cu(2+)-N bond, respectively. Due to the surface area and electrocatalytic effects of the AuNPs, the loading amount and electron transfer kinetic of the Mel-Cu(2+) 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. PMID:26931160

  15. Paper-based enzyme immobilization for flow injection electrochemical biosensor integrated with reagent-loaded cartridge toward portable modular device.

    PubMed

    Tan, Swee Ngin; Ge, Liya; Tan, Hsih Yin; Loke, Weng Keong; Gao, Jinrong; Wang, Wei

    2012-11-20

    Paper-based enzyme immobilization for a flow injection electrochemical biosensor integrated with a reagent-loaded cartridge toward a portable device was developed. A paper disk was immobilized with enzyme, then it was integrated in a flow cell as an electrochemical biosensor. A silicon tube reagent-loaded cartridge was integrated into the system, a complicated procedure was simplified as a one-click operation toward development for point-of-care applications. In this research, glucose oxidase (GOx) was employed as a model enzyme, silver ion as an inhibition reagent for GOx, and EDTA as a regeneration reagent. When GOx was inhibited by silver ions, glucose was introduced for electrochemical measurements before and after inhibited enzyme regeneration and the difference was caused by silver inhibition. The modular device has great potential for other applications, e.g., detection of enzyme activity and substrate. The platform based on double-test mode provided accurate results due to elimination of an average or control value in comparison with classical routine approaches. PMID:23116304

  16. Precise and selective sensing of DNA-DNA hybridization by graphene/Si-nanowires diode-type biosensors.

    PubMed

    Kim, Jungkil; Park, Shin-Young; Kim, Sung; Lee, Dae Hun; Kim, Ju Hwan; Kim, Jong Min; Kang, Hee; Han, Joong-Soo; Park, Jun Woo; Lee, Hosun; Choi, Suk-Ho

    2016-01-01

    Single-Si-nanowire (NW)-based DNA sensors have been recently developed, but their sensitivity is very limited because of high noise signals, originating from small source-drain current of the single Si NW. Here, we demonstrate that chemical-vapor-deposition-grown large-scale graphene/surface-modified vertical-Si-NW-arrays junctions can be utilized as diode-type biosensors for highly-sensitive and -selective detection of specific oligonucleotides. For this, a twenty-seven-base-long synthetic oligonucleotide, which is a fragment of human DENND2D promoter sequence, is first decorated as a probe on the surface of vertical Si-NW arrays, and then the complementary oligonucleotide is hybridized to the probe. This hybridization gives rise to a doping effect on the surface of Si NWs, resulting in the increase of the current in the biosensor. The current of the biosensor increases from 19 to 120% as the concentration of the target DNA varies from 0.1 to 500 nM. In contrast, such biosensing does not come into play by the use of the oligonucleotide with incompatible or mismatched sequences. Similar results are observed from photoluminescence microscopic images and spectra. The biosensors show very-uniform current changes with standard deviations ranging ~1 to ~10% by ten-times endurance tests. These results are very promising for their applications in accurate, selective, and stable biosensing. PMID:27534818

  17. Precise and selective sensing of DNA-DNA hybridization by graphene/Si-nanowires diode-type biosensors

    PubMed Central

    Kim, Jungkil; Park, Shin-Young; Kim, Sung; Lee, Dae Hun; Kim, Ju Hwan; Kim, Jong Min; Kang, Hee; Han, Joong-Soo; Park, Jun Woo; Lee, Hosun; Choi, Suk-Ho

    2016-01-01

    Single-Si-nanowire (NW)-based DNA sensors have been recently developed, but their sensitivity is very limited because of high noise signals, originating from small source-drain current of the single Si NW. Here, we demonstrate that chemical-vapor-deposition-grown large-scale graphene/surface-modified vertical-Si-NW-arrays junctions can be utilized as diode-type biosensors for highly-sensitive and -selective detection of specific oligonucleotides. For this, a twenty-seven-base-long synthetic oligonucleotide, which is a fragment of human DENND2D promoter sequence, is first decorated as a probe on the surface of vertical Si-NW arrays, and then the complementary oligonucleotide is hybridized to the probe. This hybridization gives rise to a doping effect on the surface of Si NWs, resulting in the increase of the current in the biosensor. The current of the biosensor increases from 19 to 120% as the concentration of the target DNA varies from 0.1 to 500 nM. In contrast, such biosensing does not come into play by the use of the oligonucleotide with incompatible or mismatched sequences. Similar results are observed from photoluminescence microscopic images and spectra. The biosensors show very-uniform current changes with standard deviations ranging ~1 to ~10% by ten-times endurance tests. These results are very promising for their applications in accurate, selective, and stable biosensing. PMID:27534818

  18. A Novel Bio-Sensor Based on DNA Strand Displacement

    PubMed Central

    Shi, Xiaolong; Wang, Zhiyu; Deng, Chenyan; Song, Tao; Pan, Linqiang; Chen, Zhihua

    2014-01-01

    DNA strand displacement technology performs well in sensing and programming DNA segments. In this work, we construct DNA molecular systems based on DNA strand displacement performing computation of logic gates. Specifically, a class of so-called “DNA neurons” are achieved, in which a “smart” way inspired by biological neurons encoding information is developed to encode and deliver information using DNA molecules. The “DNA neuron” is bistable, that is, it can sense DNA molecules as input signals, and release “negative” or “positive” signals DNA molecules. We design intelligent DNA molecular systems that are constructed by cascading some particularly organized “DNA neurons”, which could perform logic computation, including AND, OR, XOR logic gates, automatically. Both simulation results using visual DSD (DNA strand displacement) software and experimental results are obtained, which shows that the proposed systems can detect DNA signals with high sensitivity and accretion; moreover, the systems can process input signals automatically with complex nonlinear logic. The method proposed in this work may provide a new way to construct a sensitive molecular signal detection system with neurons spiking behavior in vitro, and can be used to develop intelligent molecular processing systems in vivo. PMID:25303242

  19. Inhibition-based first-generation electrochemical biosensors: theoretical aspects and application to 2,4-dichlorophenoxy acetic acid detection.

    PubMed

    Bollella, Paolo; Fusco, Giovanni; Tortolini, Cristina; Sanzò, Gabriella; Antiochia, Riccarda; Favero, Gabriele; Mazzei, Franco

    2016-05-01

    In this work, several theoretical aspects involved in the first-generation inhibition-based electrochemical biosensor measurements have been discussed. In particular, we have developed a theoretical-methodological approach for the characterization of the kinetic interaction between alkaline phosphatase (AlP) and 2,4-dichlorophenoxy acetic acid (2,4-D) as representative inhibitor studied by means of cyclic voltammetry and amperometry. Based on these findings, a biosensor for the fast, simple, and inexpensive determination of 2,4-D has been developed. The enzyme has been immobilized on screen-printed electrodes (SPEs). To optimize the biosensor performances, several carbon-based SPEs, namely graphite (G), graphene (GP), and multiwalled carbon nanotubes (MWCNTs), have been evaluated. AlP was immobilized on the electrode surface by means of polyvinyl alcohol with styryl-pyridinium groups (PVA-SbQ) as cross-linking agent. In the presence of ascorbate 2-phosphate (A2P) as substrate, the herbicide has been determined, thanks to its inhibition activity towards the enzyme catalyzing the oxidation of A2P to ascorbic acid (AA). Under optimum experimental conditions, the best performance in terms of catalytic efficiency has been demonstrated by MWCNTs SPE-based biosensor. The inhibition biosensor shows a linearity range towards 2,4-D within 2.1-110 ppb, a LOD of 1 ppb, and acceptable repeatability and stability. This analysis method was applied to fortified lake water samples with recoveries above 90 %. The low cost of this device and its good analytical performances suggest its application for the screening and monitoring of 2,4-D in real matrices. Graphical Abstract ᅟ. PMID:26874693

  20. Fabrication of Ultrasensitive Field-Effect Transistor DNA Biosensors by a Directional Transfer Technique Based on CVD-Grown Graphene.

    PubMed

    Zheng, Chao; Huang, Le; Zhang, Hong; Sun, Zhongyue; Zhang, Zhiyong; Zhang, Guo-Jun

    2015-08-12

    Most graphene field-effect transistor (G-FET) biosensors are fabricated through a routine process, in which graphene is transferred onto a Si/SiO2 substrate and then devices are subsequently produced by micromanufacture processes. However, such a fabrication approach can introduce contamination onto the graphene surface during the lithographic process, resulting in interference for the subsequent biosensing. In this work, we have developed a novel directional transfer technique to fabricate G-FET biosensors based on chemical-vapor-deposition- (CVD-) grown single-layer graphene (SLG) and applied this biosensor for the sensitive detection of DNA. A FET device with six individual array sensors was first fabricated, and SLG obtained by the CVD-growth method was transferred onto the sensor surface in a directional manner. Afterward, peptide nucleic acid (PNA) was covalently immobilized on the graphene surface, and DNA detection was realized by applying specific target DNA to the PNA-functionalized G-FET biosensor. The developed G-FET biosensor was able to detect target DNA at concentrations as low as 10 fM, which is 1 order of magnitude lower than those reported in a previous work. In addition, the biosensor was capable of distinguishing the complementary DNA from one-base-mismatched DNA and noncomplementary DNA. The directional transfer technique for the fabrication of G-FET biosensors is simple, and the as-constructed G-FET DNA biosensor shows ultrasensitivity and high specificity, indicating its potential application in disease diagnostics as a point-of-care tool. PMID:26203889

  1. Biosensor for label-free DNA quantification based on functionalized LPGs.

    PubMed

    Gonçalves, Helena M R; Moreira, Luis; Pereira, Leonor; Jorge, Pedro; Gouveia, Carlos; Martins-Lopes, Paula; Fernandes, José R A

    2016-10-15

    A label-free fiber optic biosensor based on a long period grating (LPG) and a basic optical interrogation scheme using off the shelf components is used for the detection of in-situ DNA hybridization. A new methodology is proposed for the determination of the spectral position of the LPG mode resonance. The experimental limit of detection obtained for the DNA was 62±2nM and the limit of quantification was 209±7nM. The sample specificity was experimentally demonstrated using DNA targets with different base mismatches relatively to the probe and was found that the system has a single base mismatch selectivity. PMID:26456729

  2. Label-free DNA Biosensor Based on SERS Molecular Sentinel on Nanowave Chip

    PubMed Central

    Ngo, Hoan Thanh; Wang, Hsin-Neng; Fales, Andrew M.; Vo-Dinh, Tuan

    2013-01-01

    Development of a rapid, cost-effective, label-free biosensor for DNA detection is important for many applications in clinical diagnosis, homeland defense, and environment monitoring. A unique label-free DNA biosensor based on Molecular Sentinel (MS) immobilized on a plasmonic ‘Nanowave’ chip, which is also referred to as a metal film over nanosphere (MFON), is presented. Its sensing mechanism is based upon the decrease of the surface-enhanced Raman scattering (SERS) intensity when Raman label tagged at one end of MS is physically separated from the MFON's surface upon DNA hybridization. This method is label-free as the target does not have to be labeled. The MFON fabrication is relatively simple and low-cost with high reproducibility based on depositing a thin shell of gold over close-packed arrays of nanospheres. The sensing process involves a single hybridization step between the DNA target sequences and the complementary MS probes on the Nanowave chip without requiring secondary hybridization or post-hybridization washing, thus resulting in rapid assay time and low reagent usage. The usefulness and potential application of the biosensor for medical diagnostics is demonstrated by detecting the human radical S-adenosyl methionine domain containing 2 (RSAD2) gene, a common inflammation biomarker. PMID:23718777

  3. Electrochemical magnetic microbeads-based biosensor for point-of-care serodiagnosis of infectious diseases.

    PubMed

    Cortina, María E; Melli, Luciano J; Roberti, Mariano; Mass, Mijal; Longinotti, Gloria; Tropea, Salvador; Lloret, Paulina; Serantes, Diego A Rey; Salomón, Francisco; Lloret, Matías; Caillava, Ana J; Restuccia, Sabrina; Altcheh, Jaime; Buscaglia, Carlos A; Malatto, Laura; Ugalde, Juan E; Fraigi, Liliana; Moina, Carlos; Ybarra, Gabriel; Ciocchini, Andrés E; Comerci, Diego J

    2016-06-15

    Access to appropriate diagnostic tools is an essential component in the evaluation and improvement of global health. Additionally, timely detection of infectious agents is critical in early diagnosis and treatment of infectious diseases. Conventional pathogen detection methods such as culturing, enzyme linked immunosorbent assay (ELISA) or polymerase chain reaction (PCR) require long assay times, and complex and expensive instruments making them not adaptable to point-of-care (PoC) needs at resource-constrained places and primary care settings. Therefore, there is an unmet need to develop portable, simple, rapid, and accurate methods for PoC detection of infections. Here, we present the development and validation of a portable, robust and inexpensive electrochemical magnetic microbeads-based biosensor (EMBIA) platform for PoC serodiagnosis of infectious diseases caused by different types of microorganisms (parasitic protozoa, bacteria and viruses). We demonstrate the potential use of the EMBIA platform for in situ diagnosis of human (Chagas disease and human brucellosis) and animal (bovine brucellosis and foot-and-mouth disease) infections clearly differentiating infected from non-infected individuals or animals. For Chagas disease, a more extensive validation of the test was performed showing that the EMBIA platform displayed an excellent diagnostic performance almost indistinguishable, in terms of specificity and sensitivity, from a fluorescent immunomagnetic assay and the conventional ELISA using the same combination of antigens. This platform technology could potentially be applicable to diagnose other infectious and non-infectious diseases as well as detection and/or quantification of biomarkers at the POC and primary care settings. PMID:26802749

  4. The development of a silica nanoparticle-based label-free DNA biosensor

    NASA Astrophysics Data System (ADS)

    Kell, Arnold J.; Pagé, Lilianne; Tan, Sophie; Charlebois, Isabelle; Boissinot, Maurice; Leclerc, Mario; Simard, Benoit

    2011-09-01

    A silica nanoparticle-based DNA biosensor capable of detecting Bacillus anthracis bacteria through the use of unlabelled ss-oligonucleotides has been developed. The biosensor makes use of the optical changes that accompany a nanoparticle-immobilized cationic conjugated polymer (polythiophene) interacting with single-stranded vs. hybridized oligonucleotides, where a fluorescence signal appears only when hybridized DNA is present (i.e. only when the ss-oligonucleotide interacting with the polymer has hybridized with its complement). In order to enhance the sensitivity of the biosensor, two different nanoparticle architectures were developed and used to elucidate how the presence of neighboring fluorophores on the nanoparticle surface affects Förster-resonant energy transfer (FRET) between the polythiophene/oligonucleotide complex (FRET donor) and the fluorophores (FRET acceptors). We demonstrate that the silica nanoparticle-based FRET platform lowers the limit of detection at least 10-fold in comparison to the polythiophene itself, and allows the detection of ~2 × 10-12 moles of ss-oligonucleotide in a 100 μL sample with a standard fluorimeter (i.e. has a limit of detection of ~2 nM ssDNA). Such nanoparticle-based biosensor platforms are beneficial because of the robustness and stability inherent to their covalent assembly and they provide a valuable new tool that may allow for the sensitive, label-free detection (the target DNA that produces the fluorescence signal is unlabelled) without the use of polymerase chain reaction.A silica nanoparticle-based DNA biosensor capable of detecting Bacillus anthracis bacteria through the use of unlabelled ss-oligonucleotides has been developed. The biosensor makes use of the optical changes that accompany a nanoparticle-immobilized cationic conjugated polymer (polythiophene) interacting with single-stranded vs. hybridized oligonucleotides, where a fluorescence signal appears only when hybridized DNA is present (i.e. only when

  5. Nanotubes, Nanowires, and Nanocantilevers in Biosensor Development

    SciTech Connect

    Wang, Jun; Liu, Guodong; Lin, Yuehe

    2007-03-08

    In this chapter, the reviews on biosensor development based on 1-D nanomaterials, CNTs, semiconducting nanowires, and some cantilevers will be introduced. The emphasis of this review will be placed on CNTs and electrochemical/electronic biosensor developments. Section 2 of this chapter gives a detailed description of carbon nanotubes-based biosensor development, from fabrication of carbon nanotubes, the strategies for construction of carbon nanotube based biosensors to their bioapplications. In the section of the applications of CNTs based biosensors, various detection principles, e. g. electrochemical, electronic, and optical method, and their applications are reviewed in detail. Section 3 introduces the method for synthesis of semiconducting nanowires, e.g. silicon nanowires, conducting polymer nanowires and metal oxide nanowires and their applications in DNA and proteins sensing. Section 4 simply describes the development for nanocantilevers based biosensors and their application in DNA and protein diagnosis. Each section starts from a brief introduction and then goes into details. Finally in the Conclusion section, the development of 1-D nanomaterials based biosensor development is summarized.

  6. Label-free detection of DNA hybridization and single point mutations in a nano-gap biosensor

    NASA Astrophysics Data System (ADS)

    Zaffino, R. L.; Mir, M.; Samitier, J.

    2014-03-01

    We describe a conductance-based biosensor that exploits DNA-mediated long-range electron transport for the label-free and direct electrical detection of DNA hybridization. This biosensor platform comprises an array of vertical nano-gap biosensors made of gold and fabricated through standard photolithography combined with focused ion beam lithography. The nano-gap walls are covalently modified with short, anti-symmetric thiolated DNA probes, which are terminated by 19 bases complementary to both the ends of a target DNA strand. The nano-gaps are separated by a distance of 50nm, which was adjusted to fit the length of the DNA target plus the DNA probes. The hybridization of the target DNA closes the gap circuit in a switch on/off fashion, in such a way that it is readily detected by an increase in the current after nano-gap closure. The nano-biosensor shows high specificity in the discrimination of base-pair mismatching and does not require signal indicators or enhancing molecules. The design of the biosensor platform is applicable for multiplexed detection in a straightforward manner. The platform is well-suited to mass production, point-of-care diagnostics, and wide-scale DNA analysis applications.

  7. A highly oriented hybrid microarray modified electrode fabricated by a template-free method for ultrasensitive electrochemical DNA recognition

    NASA Astrophysics Data System (ADS)

    Shi, Lei; Chu, Zhenyu; Dong, Xueliang; Jin, Wanqin; Dempsey, Eithne

    2013-10-01

    Highly oriented growth of a hybrid microarray was realized by a facile template-free method on gold substrates for the first time. The proposed formation mechanism involves an interfacial structure-directing force arising from self-assembled monolayers (SAMs) between gold substrates and hybrid crystals. Different SAMs and variable surface coverage of the assembled molecules play a critical role in the interfacial directing forces and influence the morphologies of hybrid films. A highly oriented hybrid microarray was formed on the highly aligned and vertical SAMs of 1,4-benzenedithiol molecules with rigid backbones, which afforded an intense structure-directing power for the oriented growth of hybrid crystals. Additionally, the density of the microarray could be adjusted by controlling the surface coverage of assembled molecules. Based on the hybrid microarray modified electrode with a large specific area (ca. 10 times its geometrical area), a label-free electrochemical DNA biosensor was constructed for the detection of an oligonucleotide fragment of the avian flu virus H5N1. The DNA biosensor displayed a significantly low detection limit of 5 pM (S/N = 3), a wide linear response from 10 pM to 10 nM, as well as excellent selectivity, good regeneration and high stability. We expect that the proposed template-free method can provide a new reference for the fabrication of a highly oriented hybrid array and the as-prepared microarray modified electrode will be a promising paradigm in constructing highly sensitive and selective biosensors.Highly oriented growth of a hybrid microarray was realized by a facile template-free method on gold substrates for the first time. The proposed formation mechanism involves an interfacial structure-directing force arising from self-assembled monolayers (SAMs) between gold substrates and hybrid crystals. Different SAMs and variable surface coverage of the assembled molecules play a critical role in the interfacial directing forces and

  8. On-Electrode Synthesis of Shape-Controlled Hierarchical Flower-Like Gold Nanostructures for Efficient Interfacial DNA Assembly and Sensitive Electrochemical Sensing of MicroRNA.

    PubMed

    Su, Shao; Wu, Yan; Zhu, Dan; Chao, Jie; Liu, Xingfen; Wan, Ying; Su, Yan; Zuo, Xiaolei; Fan, Chunhai; Wang, Lianhui

    2016-07-01

    The performance for biomolecular detection is closely associated with the interfacial structure of a biosensor, which profoundly affects both thermodynamics and kinetics of the assembly, binding and signal transduction of biomolecules. Herein, it is reported on a one-step and template-free on-electrode synthesis method for making shape-controlled gold nanostructures on indium tin oxide substrates, which provide an electrochemical sensing platform for ultrasensitive detection of nucleic acids. Thus-prepared hierarchical flower-like gold nanostructures (HFGNs) possess large surface area that can readily accommodate the assembly of DNA probes for subsequent hybridization detection. It is found that the sensitivity for electrochemical DNA sensing is critically dependent on the morphology of HFGNs. By using this new strategy, a highly sensitive electrochemical biosensor is developed for label-free detection of microRNA-21 (miRNA-21), a biomarker for lung cancers. Importantly, it is demonstrated that this biosensor can be employed to measure the miRNA-21 expression level from human lung cancer cell (A549) lysates and worked well in 100% serum, suggesting its potential for applications in clinical diagnosis and a wide range of bioanalysis. PMID:27305644

  9. Aptamer-based biosensor for label-free detection of ethanolamine by electrochemical impedance spectroscopy.

    PubMed

    Liang, Gang; Man, Yan; Jin, Xinxin; Pan, Ligang; Liu, Xinhui

    2016-09-14

    A label-free sensing assay for ethanolamine (EA) detection based on G-quadruplex-EA binding interaction is presented by using G-rich aptamer DNA (Ap-DNA) and electrochemical impedance spectroscopy (EIS). The presence of K(+) induces the Ap-DNA to form a K(+)-stabilized G-quadruplex structure which provides binding sites for EA. The sensing mechanism was further confirmed by circular dichroism (CD) spectroscopy and EIS measurement. As a result, the charge transfer resistance (RCT) is strongly increased as demonstrated by using the ferro/ferricyanide ([Fe(CN)6](3-/4-)) as a redox probe. Under the optimized conditions, a linear relationship between ΔRCT and EA concentration was obtained over the range of 0.16 nM and 16 nM EA, with a detection limit of 0.08 nM. Interference by other selected chemicals with similar structure was negligible. Analytical results of EA spiked into tap water and serum by the sensor suggested the assay could be successfully applied to real sample analysis. With the advantages of high sensitivity, selectivity and simple sensor construction, this method is potentially suitable for the on-site monitoring of EA contamination. PMID:27566359

  10. A novel mesoporous silica nanosphere matrix for the immobilization of proteins and their applications as electrochemical biosensor.

    PubMed

    Li, Juan; Qin, Xingzhang; Yang, Zhanjun; Qi, Huamei; Xu, Qin; Diao, Guowang

    2013-01-30

    A mesoporous silica nanoshpere (MSN) was proposed to modify glassy carbon electrode (GCE) for the immobilization of protein. Using glucose oxidase (GOD) as a model, direct electrochemistry of protein and biosensing at the MSN modified GCE was studied for the first time. The MNS had large surface area and offered a favorable microenvironment for facilitating the direct electron transfer between enzyme and electrode surface. Scanning electron microscopy, transmission electron microscopy, UV-vis spectroscopy and cyclic voltammetry were used to examine the interaction between GOD and the MSN matrix. The results demonstrated that the immobilized enzyme on the MSN retained its native structure and bioactivity. In addition, the electrochemical reaction showed a surface controlled, reversible two-proton and two-electron transfer process with the apparent electron transfer rate constant of 3.96 s(-1). The MNS-based glucose biosensor exhibited the two linear ranges of 0.04-2.0 mM and 2.0-4.8 mM, a high sensitivity of 14.5 mA M(-1) cm(-2) and a low detection limit of 0.02 mM at signal-to-noise of 3. The proposed biosensor showed excellent selectivity, good reproducibility, acceptable stability and could be successfully applied in the reagentless detection of glucose in real samples at -0.45 V. The work displayed that mesoporous silica nanosphere provided a promising approach for immobilizing proteins and fabrication of excellent biosensors. PMID:23597897

  11. Direct electron transfer glucose biosensor based on glucose oxidase self-assembled on electrochemically reduced carboxyl graphene.

    PubMed

    Liang, Bo; Fang, Lu; Yang, Guang; Hu, Yichuan; Guo, Xishan; Ye, Xuesong

    2013-05-15

    A glucose biosensor based on direct electron transfer of glucose oxidase (GOD) self-assembled on the surface of the electrochemically reduced carboxyl graphene (ERCGr) modified glassy carbon electrode has been reported. X-ray photoelectron spectroscopy (XPS) analyses of ERCGr indicate most of the oxygen-containing groups such as epoxy/ether groups and hydroxyl groups in the carboxyl graphene were eliminated, while carboxylic acid groups remained. GOD was immobilized on the ERCGr modified glassy carbon electrode via self-assembly. The cyclic voltammetric result of the electrode shows a pair of well-defined and quasi-reversible redox peaks with a formal potential of -0.467 V and a peak to peak separation of 49 mV, revealing that the direct electron transfer between GOD and the electrode has been achieved. The proposed biosensor exhibits a linear response to glucose concentrations ranging from 2 to 18 mM with a detection limit of 0.02 mM. Moreover, this facile, fast, environment-friendly and economical preparation strategy of ERCGr may be extended for the preparation of other graphene based enzyme electrode biosensors. PMID:23298623

  12. Scanning Electrochemical Microscopy of DNA Monolayers Modified with Nile Blue

    PubMed Central

    Gorodetsky, Alon A.; Hammond, William J.; Hill, Michael G.; Slowinski, Krzysztof; Barton, Jacqueline K.

    2009-01-01

    Scanning electrochemical microscopy (SECM) is used to probe long-range charge transport (CT) through DNA monolayers containing the redox-active Nile Blue (NB) intercalator covalently affixed at a specific location in the DNA film. At substrate potentials negative of the formal potential of covalently attached NB, the electrocatalytic reduction of Fe(CN)63− generated at the SECM tip is observed only when NB is located at the DNA/solution interface; for DNA films containing NB in close proximity to the DNA/electrode interface, the electrocatalytic effect is absent. This behavior is consistent with both rapid DNA-mediated CT between the NB intercalator and the gold electrode as well as a rate-limiting electron transfer between NB and the solution phase Fe(CN)63−. The DNA-mediated nature of the catalytic cycle is confirmed through sequence-specific and localized detection of attomoles of TATA-binding protein, a transcription factor that severely distorts DNA upon binding. Importantly, the strategy outlined here is general and allows for the local investigation of the surface characteristics of DNA monolayers both in the absence and in the presence of DNA binding proteins. These experiments highlight the utility of DNA-modified electrodes as versatile platforms for SECM detection schemes that take advantage of CT mediated by the DNA base pair stack. PMID:19053641

  13. CdS/MoS2 heterojunction-based photoelectrochemical DNA biosensor via enhanced chemiluminescence excitation.

    PubMed

    Zang, Yang; Lei, Jianping; Hao, Qing; Ju, Huangxian

    2016-03-15

    This work developed a CdS/MoS2 heterojunction-based photoelectrochemical biosensor for sensitive detection of DNA under the enhanced chemiluminescence excitation of luminol catalyzed by hemin-DNA complex. The CdS/MoS2 photocathode was prepared by the stepwise assembly of MoS2 and CdS quantum dots (QDs) on indium tin oxide (ITO), and achieved about 280% increasing of photocurrent compared to pure CdS QDs electrode due to the formation of heterostructure. High photoconversion efficiency in the photoelectrochemical system was identified to be the rapid spatial charge separation of electron-hole pairs by the extension of electron transport time and electron lifetime. In the presence of target DNA, the catalytic hairpin assembly was triggered, and simultaneously the dual hemin-labeled DNA probe was introduced to capture DNA/CdS/MoS2 modified ITO electrode. Thus the chemiluminescence emission of luminol was enhanced via hemin-induced mimetic catalysis, leading to the physical light-free photoelectrochemical strategy. Under optimized conditions, the resulting photoelectrode was proportional to the logarithm of target DNA concentration in the range from 1 fM to 100 pM with a detection limit of 0.39 fM. Moreover, the cascade amplification biosensor demonstrated high selectivity, desirable stability and good reproducibility, showing great prospect in molecular diagnosis and bioanalysis. PMID:26476013

  14. An electrochemical biosensor with nanointerface for lactate detection based on lactate dehydrogenase immobilized on zinc oxide nanorods.

    PubMed

    Nesakumar, Noel; Thandavan, Kavitha; Sethuraman, Swaminathan; Krishnan, Uma Maheswari; Rayappan, John Bosco Balaguru

    2014-01-15

    Hepatic immaturity is observed particularly in children whose age is under three, when the lactate concentration is greater than the normal level in blood. An electrochemical lactate biosensor was developed by immobilizing lactate dehydrogenase (LDH) on to ZnO nanorods at pH 7.4 via chitosan. Growth of polycrystalline ZnO nanorods towards (101) plane was confirmed using XRD. The FE-SEM study revealed the formation of ZnO nanorods with an aspect ratio of 3.24. Immobilization of LDH on ZnO nanorods was confirmed using FTIR spectra and surface coverage. Electrochemical studies were carried out through cyclic voltammetry and amperometry using three electrode system with Au/NanoZnO/LDH as working electrode, Ag/AgCl in 0.1 M KCl as reference electrode and Pt wire as counter electrode. The sensitivity of the biosensor was found to be 1.832 μA μmol(-1) L exhibiting linearity 0.2-0.8 μmol L(-1) with the detection and quantification limits of 4.73 and 15.75 nmol L(-1) respectively. The response time of Au/NanoZnO/LDH bioelectrode was found to be <1 s. Prediction band for net current was framed to enhance specificity. Michaelis-Menten constant (KM(app)) and maximum rate (Imax) values for immobilized LDH were found to be 0.38 μmol L(-1) and 2.798 μA respectively. Repeatability and reproducibility of LDH biosensor were also reported. PMID:24231089

  15. A highly sensitive electrochemical biosensor for catechol using conducting polymer reduced graphene oxide-metal oxide enzyme modified electrode.

    PubMed

    Sethuraman, V; Muthuraja, P; Anandha Raj, J; Manisankar, P

    2016-10-15

    The fabrication, characterization and analytical performances were investigated for a catechol biosensor, based on the PEDOT-rGO-Fe2O3-PPO composite modified glassy carbon (GC) electrode. The graphene oxide (GO) doped conducting polymer poly (3,4-ethylenedioxythiophene) (PEDOT) was prepared through electrochemical polymerization by potential cycling. Reduction of PEDOT-GO was carried out by amperometric method. Fe2O3 nanoparticles were synthesized in ethanol by hydrothermal method. The mixture of Fe2O3, PPO and glutaraldehyde was casted on the PEDOT-rGO electrode. The surface morphology of the modified electrodes was studied by FE-SEM and AFM. Cyclic voltammetric studies of catechol on the enzyme modified electrode revealed higher reduction peak current. Determination of catechol was carried out successfully by Differential Pulse Voltammetry (DPV) technique. The fabricated biosensor investigated shows a maximum current response at pH 6.5. The catechol biosensor exhibited wide sensing linear range from 4×10(-8) to 6.20×10(-5)M, lower detection limit of 7×10(-9)M, current maxima (Imax) of 92.55µA and Michaelis-Menten (Km) constant of 30.48µM. The activation energy (Ea) of enzyme electrode is 35.93KJmol(-1) at 50°C. There is no interference from d-glucose and l-glutamic acid, ascorbic acid and o-nitrophenol. The PEDOT-rGO-Fe2O3-PPO biosensor was stable for at least 75 days when stored in a buffer at about 4°C. PMID:26751827

  16. Detection of Non-PCR Amplified S. enteritidis Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study

    PubMed Central

    Vetrone, Sylvia A.; Huarng, Michael C.; Alocilja, Evangelyn C.

    2012-01-01

    Bacterial pathogens pose an increasing food safety and bioterrorism concern. Current DNA detection methods utilizing sensitive nanotechnology and biosensors have shown excellent detection, but require expensive and time-consuming polymerase chain reaction (PCR) to amplify DNA targets; thus, a faster, more economical method is still essential. In this proof-of-concept study, we investigated the ability of a gold nanoparticle-DNA (AuNP-DNA) biosensor to detect non-PCR amplified genomic Salmonella enterica serovar Enteritidis (S. enteritidis) DNA, from pure or mixed bacterial culture and spiked liquid matrices. Non-PCR amplified DNA was hybridized into sandwich-like structures (magnetic nanoparticles/DNA/AuNPs) and analyzed through detection of gold voltammetric peaks using differential pulse voltammetry. Our preliminary data indicate that non-PCR amplified genomic DNA can be detected at a concentration as low as 100 ng/mL from bacterial cultures and spiked liquid matrices, similar to reported PCR amplified detection levels. These findings also suggest that AuNP-DNA biosensors are a first step towards a viable detection method of bacterial pathogens, in particular, for resource-limited settings, such as field-based or economically limited conditions. Future efforts will focus on further optimization of the DNA extraction method and AuNP-biosensors, to increase sensitivity at lower DNA target concentrations from food matrices comparable to PCR amplified DNA detection strategies. PMID:23112611

  17. Electrochemical DNA sensor for specific detection of picomolar Hg(II) based on exonuclease III-assisted recycling signal amplification.

    PubMed

    Gan, Xiaorong; Zhao, Huimin; Chen, Shuo; Quan, Xie

    2015-03-21

    An ultrasensitive methodology was successfully developed for the quantitative detection of picomolar Hg(2+) based on the combination of thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination chemistry and exonuclease III-aided recycling signal amplification. Single-strand probe DNA was immobilized on an Au electrode via an Au-S bond. In the presence of Hg(2+), the probe DNA hybridized with the target DNA containing four thymine-thymine (T-T) mismatches via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. Then the probe DNA in the DNA duplex was specifically recognized and selectively digested by exonuclease III; in contrast the target DNA was safely dissociated from the DNA duplexes to subsequently hybridize with a new signal probe, leading to target recycling and signal amplification. As a result, the peak current caused by the electrostatic interactions of [Ru(NH3)6](3+) cations with the backbone of the probe DNA decreased by different degrees, corresponding to the Hg(2+) concentrations. Under the optimum conditions, the proposed electrochemical DNA biosensor showed a robust detection limit as low as 1 pM (S/N = 3), with a wide linear range from 0.01 to 500 nM and good selectivity. In addition, the proposed method was successfully applied to assay Hg(2+) in real environmental samples. PMID:25676090

  18. Electrochemical Sensors Based on Organic Conjugated Polymers

    PubMed Central

    Rahman, Md. Aminur; Kumar, Pankaj; Park, Deog-Su; Shim, Yoon-Bo

    2008-01-01

    Organic conjugated polymers (conducting polymers) have emerged as potential candidates for electrochemical sensors. Due to their straightforward preparation methods, unique properties, and stability in air, conducting polymers have been applied to energy storage, electrochemical devices, memory devices, chemical sensors, and electrocatalysts. Conducting polymers are also known to be compatible with biological molecules in a neutral aqueous solution. Thus, these are extensively used in the fabrication of accurate, fast, and inexpensive devices, such as biosensors and chemical sensors in the medical diagnostic laboratories. Conducting polymer-based electrochemical sensors and biosensors play an important role in the improvement of public health and environment because rapid detection, high sensitivity, small size, and specificity are achievable for environmental monitoring and clinical diagnostics. In this review, we summarized the recent advances in conducting polymer-based electrochemical sensors, which covers chemical sensors (potentiometric, voltammetric, amperometric) and biosensors (enzyme based biosensors, immunosensors, DNA sensors).

  19. Electrochemical Sensors Based on Carbon Nanotubes

    PubMed Central

    Saleh Ahammad, A. J.; Lee, Jae-Joon; Rahman, Md. Aminur

    2009-01-01

    This review focuses on recent contributions in the development of the electrochemical sensors based on carbon nanotubes (CNTs). CNTs have unique mechanical and electronic properties, combined with chemical stability, and behave electrically as a metal or semiconductor, depending on their structure. For sensing applications, CNTs have many advantages such as small size with larger surface area, excellent electron transfer promoting ability when used as electrodes modifier in electrochemical reactions, and easy protein immobilization with retention of its activity for potential biosensors. CNTs play an important role in the performance of electrochemical biosensors, immunosensors, and DNA biosensors. Various methods have been developed for the design of sensors using CNTs in recent years. Herein we summarize the applications of CNTs in the construction of electrochemical sensors and biosensors along with other nanomaterials and conducting polymers. PMID:22574013

  20. Detection of DNA and poly-l-lysine using CVD graphene-channel FET biosensors

    NASA Astrophysics Data System (ADS)

    Kakatkar, Aniket; Abhilash, T. S.; De Alba, R.; Parpia, J. M.; Craighead, H. G.

    2015-03-01

    A graphene channel field-effect biosensor is demonstrated for detecting the binding of double-stranded DNA and poly-l-lysine. Sensors consist of chemical vapor deposition graphene transferred using a clean, etchant-free transfer method. The presence of DNA and poly-l-lysine are detected by the conductance change of the graphene transistor. A readily measured shift in the Dirac voltage (the voltage at which the graphene’s resistance peaks) is observed after the graphene channel is exposed to solutions containing DNA or poly-l-lysine. The ‘Dirac voltage shift’ is attributed to the binding/unbinding of charged molecules on the graphene surface. The polarity of the response changes to positive direction with poly-l-lysine and negative direction with DNA. This response results in detection limits of 8 pM for 48.5 kbp DNA and 11 pM for poly-l-lysine. The biosensors are easy to fabricate, reusable and are promising as sensors of a wide variety of charged biomolecules.

  1. A novel and simple cell-based electrochemical impedance biosensor for evaluating the combined toxicity of DON and ZEN.

    PubMed

    Gu, Wenshu; Zhu, Pei; Jiang, Donglei; He, Xingxing; Li, Yun; Ji, Jian; Zhang, Lijuan; Sun, Yange; Sun, Xiulan

    2015-08-15

    In this study, a novel and simple cell-based electrochemical biosensor was developed to assess the individual and combined toxicity of deoxynivalenol (DON) and zearalenone (ZEN) on BEL-7402 cells. The sensor was fabricated by modification with AuNPs, p-aminothiophenol, and folic acid in succession. The BEL-7402 cells which had a good activity were adhered on the electrode through the high affinity between the folate receptor and folic acid selectivity. We used the collagen to maintain the cell adhesion and viability. Electrochemical impedance spectroscopy (EIS) was developed to evaluate the individual and combined toxicity of DON and ZEN. Our results indicate that DON and ZEN caused a marked decrease in the cell viability in a dose-dependent manner. The value of electrochemical impedance spectroscopy decreased with the concentration of DON and ZEN in range of 0.1-20, 0.1-50 μg/ml with the detection limit as 0.03, 0.05 μg/ml, respectively, the IC50 for DON and ZEN as obtained by the proposed electrochemical method were 7.1 μg/ml and 24.6 μg/ml, respectively, and the combination of two mycotoxins appears to generate an additive response. The electrochemical cytotoxicity evaluation result was confirmed by biological assays. Compared to conventional methods, this electrochemical test is inexpensive, highly sensitive, and fast to respond, with long-term monitoring and real-time measurements. The proposed method provides a new avenue for evaluating the toxicity of mycotoxins. PMID:25863342

  2. An InN/InGaN Quantum Dot Electrochemical Biosensor for Clinical Diagnosis

    PubMed Central

    Alvi, Naveed ul Hassan; Gómez, Victor J.; Rodriguez, Paul E.D. Soto; Kumar, Praveen; Zaman, Saima; Willander, Magnus; Nötzel, Richard

    2013-01-01

    Low-dimensional InN/InGaN quantum dots (QDs) are demonstrated for realizing highly sensitive and efficient potentiometric biosensors owing to their unique electronic properties. The InN QDs are biochemically functionalized. The fabricated biosensor exhibits high sensitivity of 97 mV/decade with fast output response within two seconds for the detection of cholesterol in the logarithmic concentration range of 1 × 10−6 M to 1 × 10−3 M. The selectivity and reusability of the biosensor are excellent and it shows negligible response to common interferents such as uric acid and ascorbic acid. We also compare the biosensing properties of the InN QDs with those of an InN thin film having the same surface properties, i.e., high density of surface donor states, but different morphology and electronic properties. The sensitivity of the InN QDs-based biosensor is twice that of the InN thin film-based biosensor, the EMF is three times larger, and the response time is five times shorter. A bare InGaN layer does not produce a stable response. Hence, the superior biosensing properties of the InN QDs are governed by their unique surface properties together with the zero-dimensional electronic properties. Altogether, the InN QDs-based biosensor reveals great potential for clinical diagnosis applications. PMID:24132228

  3. Cell-Based Biosensor to Report DNA Damage in Micro- and Nanosystems

    PubMed Central

    2015-01-01

    Understanding how newly engineered micro- and nanoscale materials and systems that interact with cells impact cell physiology is crucial for the development and ultimate adoption of such technologies. Reports regarding the genotoxic impact of forces applied to cells in such systems that can both directly or indirectly damage DNA emphasize the need for developing facile methods to assess how materials and technologies affect cell physiology. To address this need we have developed a TurboRFP-based DNA damage reporter cell line in NIH-3T3 cells that fluoresce to report genotoxic stress caused by a wide variety of agents, from chemical genotoxic agents to UV-C radiation. Our biosensor was successfully implemented in reporting the genotoxic impact of nanomaterials, demonstrating the ability to assess size dependent geno- and cyto-toxicity. The biosensor cells can be assayed in a high throughput, noninvasive manner, with no need for overly sophisticated equipment or additional reagents. We believe that this open-source biosensor is an important resource for the community of micro- and nanomaterials and systems designers and users who wish to evaluate the impact of systems and materials on cell physiology. PMID:25001406

  4. Effective immobilization of DNA for development of polypyrrole nanowires based biosensor

    NASA Astrophysics Data System (ADS)

    Tran, Thi Luyen; Chu, Thi Xuan; Huynh, Dang Chinh; Pham, Duc Thanh; Luu, Thi Hoai Thuong; Mai, Anh Tuan

    2014-09-01

    This paper reports an easy technique for immobilization of the DNA to the conducting polymer polypyrrole nanowires (PPy NWs). The nanowires were electrochemically synthesized on the surface of working electrode in the presence of gelatin as a soft mold. The structure of obtained PPy NWs was investigated by Scanning Electron Microscopy (SEM), Fourier Transform Infrared (FTIR) spectroscopy and Surface Enhanced Raman Spectroscopy (SERS). The DNA strands were directly immobilized on the PPy NWs. The amino groups at the up-end of the PPy nanowires facilitate the linkage with the phosphate groups of the probe DNA. The DNA immobilization and hybridization were characterized by Electrochemical Impedance Spectroscopy (EIS). The initial results show that the sensor responses to 10 pM of DNA sequence in the solution.

  5. Exonuclease III-aided autocatalytic DNA biosensing platform for immobilization-free and ultrasensitive electrochemical detection of nucleic acid and protein.

    PubMed

    Liu, Shufeng; Lin, Ying; Wang, Li; Liu, Tao; Cheng, Chuanbin; Wei, Wenji; Tang, Bo

    2014-04-15

    Homogenous electrochemical biosensor has attracted substantial attention owing to its simplicity, rapid response, and improved recognition efficiency compared with heterogeneous biosensor, but the relatively low detection sensitivity and the limited detection analytes prohibit its potential applications. To address these issues, herein, a simple, rapid, isothermal, and ultrasensitive homogeneous electrochemical DNA biosensing platform for target DNA and protein detection has been developed on the basis of an exonuclease III (Exo III)-aided autocatalytic target recycling strategy. A ferrocene-labeled hairpin probe (HP1) is ingeniously designed, which contains a protruding DNA fragment at 3'-termini as the recognition unit for target DNA. Also, the DNA fragment that could be used as secondary target analogue was introduced, but it was caged in the stem region of HP1. In the presence of target DNA, its recognition with the protruding fragment of HP1 triggered the Exo III cleavage process, accompanied with the target recycling and autonomous generation of secondary target analogues. This accordingly resulted into the autonomous accumulation of ferrocene-labeled mononucleotide, inducing a distinct increase in the electrochemical signal owing to its elevated diffusivity toward indium tin oxide (ITO) electrode surface. The autocatalytic biosensing system was further extended for protein detection by advising an aptamer hairpin switch with the use of thrombin as a model analyte. The current developed autocatalytic and homogeneous strategy provided an ultrasensitive electrochemical detection of DNA and thrombin down to the 0.1 and 5 pM level, respectively, with a high selectivity. It should be further used as a general autocatalytic and homogeneous strategy toward the detection of a wide spectrum of analytes and may be associated with more analytical techniques. Thus, it holds great potential for the development of ultrasensitive biosensing platform for the applications in

  6. Integrated biochip for PCR-based DNA amplification and detection on capacitive biosensors

    NASA Astrophysics Data System (ADS)

    Moschou, D.; Vourdas, N.; Filippidou, M. K.; Tsouti, V.; Kokkoris, G.; Tsekenis, G.; Zergioti, I.; Chatzandroulis, S.; Tserepi, A.

    2013-05-01

    Responding to an increasing demand for LoC devices to perform bioanalytical protocols for disease diagnostics, the development of an integrated LoC device consisting of a μPCR module integrated with resistive microheaters and a biosensor array for disease diagnostics is presented. The LoC is built on a Printed Circuit Board (PCB) platform, implementing both the amplification of DNA samples and DNA detection/identification on-chip. The resistive microheaters for PCR and the wirings for the sensor read-out are fabricated by means of standard PCB technology. The microfluidic network is continuous-flow, designed to perform 30 PCR cycles with heated zones at constant temperatures, and is built onto the PCB utilizing commercial photopatternable polyimide layers. Following DNA amplification, the product is driven in a chamber where a Si-based biosensor array is placed for DNA detection through hybridization. The sensor array is tested for the detection of mutations of the KRAS gene, responsible for colon cancer.

  7. Application of hydrophobic palladium nanoparticles for the development of electrochemical glucose biosensor.

    PubMed

    Li, Zhongping; Wang, Xu; Wen, Guangming; Shuang, Shaomin; Dong, Chuan; Paau, Man Chin; Choi, Martin M F

    2011-07-15

    An amperometric glucose biosensor based on an n-alkylamine-stabilized palladium nanoparticles (PdNPs)-glucose oxidase (GOx) modified glassy carbon (GC) electrode has been successfully fabricated. PdNPs were initially synthesized by a biphase mixture of water and toluene method using n-alkylamines (dodecylamine, C₁₂-NH₂ and octadecylamine, C₁₈-NH₂) as stabilizing ligands. The performance of the PdNPs-GOx/GC biosensor was studied by cyclic voltammetry. The optimum working potential for amperometric measurement of glucose in pH 7.0 phosphate buffer solution is -0.02 V (vs. Ag/AgCl). The analytical performance of the biosensor prepared from C₁₈-PdNPs-GOx is better than that of C₁₂-PdNPs-GOx. The C₁₈-PdNPs-GOx/GC biosensor exhibits a fast response time of ca. 3s, a detection limit of 3.0 μM (S/N=3) and a linear range of 3.0 μM-8.0 mM. The linear dependence of current density with glucose concentration is 70.8 μA cm⁻² mM⁻¹. The biosensor shows good stability, repeatability and reproducibility. It has been successfully applied to determine the glucose content in human blood serum samples. PMID:21612909

  8. Electrochemical discrimination between G-quadruplex and duplex DNA.

    PubMed

    De Rache, Aurore; Doneux, Thomas; Buess-Herman, Claudine

    2014-08-19

    Analytical tools enabling the discrimination between duplex DNA and G-quadruplex DNA are necessary to unravel the biological function(s) of G-quadruplexes. A methodology relying on the electrochemical response of the electroactive hexaammineruthenium(III) cation at DNA-modified surfaces is presented. A characteristic voltammetric peak is evidenced for all the investigated G-quadruplex sequences, encompassing various types of folding and numbers of quartets. In contrast, no such peak is detected for dsDNA sequences. The occurrence of the voltammetric peak is the consequence of a strong association between the hexaammineruthenium ligand and the surface-immobilized G-quadruplexes. The peak potential points to a significant contribution of nonelectrostatic interactions between the electroactive ligand and G-quadruplexes. The very good efficiency of the discrimination methodology is demonstrated by comparing a G-quadruplex and its corresponding duplex. PMID:25048277

  9. Determination of Organophosphate Pesticides at a Carbon Nanotube/Organophosphorus Hydrolase Electrochemical Biosensor

    SciTech Connect

    Deo, R P.; Wang, Joseph; Block, I; Mulchandani, Ashok; Joshi, K; Trojanowicz, M; Scholz, F; Chen, Wilfred; Lin, Yuehe

    2005-02-08

    An amperometric biosensor for organophosphorus (OP) pesticides based on a carbon-nanotube (CNT) modified transducer and an organophosphorus hydrolase (OPH) biocatalyst is described. A bilayer approach with the OPH layer atop of the CNT film was used for preparing the CNT/OPH biosensor. The CNT layer leads to a greatly improved anodic detection of the enzymatically-generated p-nitrophenol product, including higher sensitivity and stability. The sensor performance was optimized with respect to the surface modification and operating conditions. Under the optimal conditions the biosensor was used to measure as low as 0.15 {micro}M paraoxon and 0.8 {micro}M methyl parathion with sensitivities of 25 and 6 nA/{micro}M, respectively.

  10. Application of photochemical reaction in electrochemical detection of DNA intercalation.

    PubMed

    Pandey, P C; Weetall, H H

    1994-04-15

    A flow injection analysis (FIA) system for the detection of the compounds that intercalate within DNA is reported. A derivative of 9,10-anthraquinone has been used as the reference compound for photoelectrochemical detection. The sodium salts of 9,10-anthraquinone-2,6-disulfonic acid and 9,10-anthraquinone-2-sulfonic acid are photochemically activated and then reduced in the presence of an electron donor (glucose). The electrochemical signal is based on the measurement of the anodic current resulting from the oxidation of the reduced form of 9,10-anthraquinone. The reduced form of the 9,10-anthraquinone is oxidized through a mediated mechanism at the surface of a tetracyanoquinodimethane-(TCNQ)-modified graphite paste electrode covered by a Nucleopore membrane. TCNQ acts as an efficient mediator for the oxidation of reduced 9,10-anthraquinone. Cyclic voltammetry, photocyclic voltammetry, and the photoelectrochemical FIA response of 9,10-anthraquinone are reported. Experimental results show that these anthraquinones can be intercalated within the helix of double-stranded calf thymus DNA. The anthraquinone molecules that are intercalated within DNA cannot be oxidized due to their limited transport to the modified electrode surface. This results in a decrease in the anodic current at a constant concentration of anthraquinone after intercalation. There is a linear relation between the decrease in electrochemical response and the DNA concentration at a constant concentration of anthraquinone. The intercalated anthraquinone molecules can be completely replaced by another intercalating agent (i.e., ethidium bromide) that is more strongly intercalated within DNA, thereby regenerating the electrochemical response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8210041

  11. An upconversion fluorescent resonant energy transfer biosensor for hepatitis B virus (HBV) DNA hybridization detection.

    PubMed

    Zhu, Hao; Lu, Feng; Wu, Xing-Cai; Zhu, Jun-Jie

    2015-11-21

    A novel fluorescent resonant energy transfer (FRET) biosensor was fabricated for the detection of hepatitis B virus (HBV) DNA using poly(ethylenimine) (PEI) modified upconversion nanoparticles (NH2-UCNPs) as energy donor and gold nanoparticles (Au NPs) as acceptor. The PEI modified upconversion nanoparticles were prepared directly with a simple one-pot hydrothermal method, which provides high quality amino-group functionalized UCNPs with uniform morphology and strong upconversion luminescence. Two single-stranded DNA strands, which were partially complementary to each other, were then conjugated with NH2-UCNPs and Au NPs. When DNA conjugated NH2-UCNPs and Au NPs are mixed together, the hybridization between complementary DNA sequences on UCNPs and Au NPs will lead to the quenching of the upconversion luminescence due to the FRET process. Meanwhile, upon the addition of target DNA, Au NPs will leave the surface of the UCNPs and the upconversion luminescence can be restored because of the formation of the more stable double-stranded DNA on the UCNPs. The sensor we fabricated here for target DNA detection shows good sensitivity and high selectivity, which has the potential for clinical applications in the analysis of HBV and other DNA sequences. PMID:26421323

  12. Development of a biosensor for electrochemical detection of tumor-associated proteins in blood plasma of cancer patients by aptamers.

    PubMed

    Zamay, G S; Zamay, T N; Kolovskaya, O S; Krat, A V; Glazyrin, Yu E; Dubinina, A V; Zamay, A S

    2016-01-01

    A molecular biosensor based on DNA aptamers (aptasensor) for the diagnosis of lung cancer in blood plasma samples was designed. To create the aptasensor, the aptamer 17_80, obtained in the study of postoperative material, was used. The affinity and binding selectivity of the aptamer 17_80 to the lung tumor tissue was confirmed on histological sections of postmortem samples of lung tissue. Using affinity enrichment and mass spectrometry, a possible target molecule of the aptamer 17_80, vimentin, was found. PMID:27025492

  13. An Analytical Model for Thermal Effect of Microcantilever-DNA Biosensors

    NASA Astrophysics Data System (ADS)

    Tan, Zou-Qing; Zhang, Neng-Hui

    2013-06-01

    The thermal effect of microcantilever-DNA biosensors is investigated by the energy method. Based on a liquid crystal theory for DNA solutions and a two-variable method for laminated cantilevers, an analytical model for nanomechanical cantilever motion under the combination of bio-interactions and thermal loadings is provided and then it is extended to T-shaped cantilevers. Then, the effects of chemo-physical properties of DNA biofilm (i.e., grafting density, nucleotide number, and ionic strength) and temperature change on deflections are discussed. In order to reduce noise signals, the controlling temperature and size optimization of cantilevers with different substrate materials and ionic strengths are also studied. Results show that SU-8 polymer cantilevers can preserve the sensitivity of molecule adsorption and thermal stability, which agrees well with the related experiments; the layer-to-layer thickness ratio of SU-8 polymer cantilevers should be as small as possible, while for silicon nitride cantilevers, there exists an optimal value. These results help to understand the sensitivity and reproducibility of biosensors.

  14. Fabrication of a facile electrochemical biosensor for hydrogen peroxide using efficient catalysis of hemoglobin on the porous Pd@Fe3O4-MWCNT nanocomposite.

    PubMed

    Baghayeri, Mehdi; Veisi, Hojat

    2015-12-15

    In this work, a sensitive amperometric biosensor for hydrogen peroxide based on synergetic catalysis of hemoglobin and porous Pd@Fe3O4-MWCNT nanocomposite has been constructed. With attention to the utilities of large surface area and outstanding catalytic performance, Pd@Fe3O4-MWCNT nanocomposite was employed as the nano-stabilizer for the immobilization of hemoglobin (Hb). The immobilized Hb on the surface of nanocomposite as an electrochemical biosensor efficiently catalyzed the reduction of hydrogen peroxide, amplified the electrochemical signal and enhanced the sensitivity. Results of voltammetry and electrochemical impedance examinations showed that the nanocomposite could enhance the electron conductivity and provide more sites for the immobilization of Hb. A linear response from 0.2-500 µM with detection limit of 0.063 µM for hydrogen peroxide was achieved. The apparent Michaelis-Menten constant Kapp(M) value was 21 µM. Thus, the nanocomposite could be applied for fabrication of a third generation biosensor for hydrogen peroxide with high sensitivity, selectivity and low detection limit. The excellent performance of the biosensor indicated its promising prospect as a valuable tool in simple and fast hydrogen peroxide detection in environmental and clinical applications. PMID:26143458

  15. Stable and Reusable Electrochemical Biosensor for Poly(ADP-ribose) Polymerase and Its Inhibitor Based on Enzyme-Initiated Auto-PARylation.

    PubMed

    Xu, Yuanyuan; Liu, Li; Wang, Zhaoyin; Dai, Zhihui

    2016-07-27

    A stable and reusable electrochemical biosensor for the label-free detection of poly(ADP-ribose) polymerase (PARP) is designed in this work. C-kit-1, a thiol-modified G-quadruplex oligonucleotide, is first self-assembled on a gold electrode surface. The G-quadruplex structure of c-kit-1 can specifically tether and activate PARP, resulting in the generation of negatively charged poly(ADP-ribose) polymer (PAR). On the basis of electrostatic attraction, PAR facilitates the surface accumulation of positively charged electrochemical signal molecules. Through the characterization of electrochemical signal molecules, the label-free quantification of PARP is simply implemented. On the basis of the proposed method, selective quantification of PARP can be achieved over the linear range from 0.01 to 1 U with a calculated detection limit of 0.003U. Further studies also demonstrate the applicability of the proposed method to biosamples revealing the broad potential in practical applications. Furthermore, inhibitor of PARP has also been detected with this biosensor. Meanwhile, benefited from self-assembly on solid surface, this biosensor possesses two important features, i.e., reusability and stability, which are desirable in related biosensors. PMID:27367274

  16. An intimately bonded titanate nanotube-polyaniline-gold nanoparticle ternary composite as a scaffold for electrochemical enzyme biosensors.

    PubMed

    Liu, Xiaoqiang; Zhu, Jie; Huo, Xiaohe; Yan, Rui; Wong, Danny K Y

    2016-03-10

    In this work, titanate nanotubes (TNTs), polyaniline (PANI) and gold nanoparticles (GNPs) were assembled to form a ternary composite, which was then applied on an electrode as a scaffold of an electrochemical enzyme biosensor. The scaffold was constructed by oxidatively polymerising aniline to produce an emeraldine salt of PANI on TNTs, followed by gold nanoparticle deposition. A novel aspect of this scaffold lies in the use of the emeraldine salt of PANI as a molecular wire between TNTs and GNPs. Using horseradish peroxidase (HRP) as a model enzyme, voltammetric results demonstrated that direct electron transfer of HRP was achieved at both TNT-PANI and TNT-PANI-GNP-modified electrodes. More significantly, the catalytic reduction current of H2O2 by HRP was ∼75% enhanced at the TNT-PANI-GNP-modified electrode, compared to that at the TNT-PANI-modified electrode. The heterogeneous electron transfer rate constant of HRP was found to be ∼3 times larger at the TNT-PANI-GNP-modified electrode than that at the TNT-PANI-modified electrode. Based on chronoamperometric detection of H2O2, a linear range from 1 to 1200 μM, a sensitivity of 22.7 μA mM(-1) and a detection limit of 0.13 μM were obtained at the TNT-PANI-GNP-modified electrode. The performance of the biosensor can be ascribed to the superior synergistic properties of the ternary composite. PMID:26893086

  17. Electrochemical and electrophoretic deposition of enzymes: principles, differences and application in miniaturized biosensor and biofuel cell electrodes.

    PubMed

    Ammam, Malika

    2014-08-15

    Recent advances in nano-biotechnology have made it possible to realize a great variety of enzyme electrodes suitable for sensing and energy applications. In coating miniaturized electrodes with enzymes, there is no doubt that most of the available deposition processes suffer from the difficulty in depositing uniform and reproducible coatings of the active enzyme on the miniature transducer element. This mini-review highlights the promising prospects of two techniques, electrochemical deposition (ECD) and electrophoretic deposition (EPD), in enzyme immobilization onto miniaturized electrodes and their use as biosensors and biofuel cells. The main differences between ECD and EPD are described and highlighted in the sense to make it clear to the reader that both techniques employ electric fields to deposit enzyme but the conditions from which each process is achieved and hence the mechanisms are quite different. Many aspects dealing with deposition of enzyme under ECD and EPD are considered including surface charge of enzyme, its migration under the applied electric field and its precipitation on the electrode. Still all issues discussed in this mini-review are generic and need to be followed in the future by extensive theoretical and experimental research analysis. Finally, the advantages of ECD and EPD in fabrication of miniature biosensor and biofuel cell electrodes are described and discussed. PMID:24632138

  18. Invertase inhibition based electrochemical sensor for the detection of heavy metal ions in aqueous system: Application of ultra-microelectrode to enhance sucrose biosensor's sensitivity.

    PubMed

    Bagal-Kestwal, Dipali; Karve, Meena S; Kakade, Bhalchandra; Pillai, Vijayamohanan K

    2008-12-01

    We are reporting fabrication and characterization of electrochemical sucrose biosensor using ultra-microelectrode (UME) for the detection of heavy metal ions (Hg(II), Ag(I), Pb(II) and Cd(II)). The working UME, with 25 microm diameter, was modified with invertase (INV, EC: 3.2.1.26) and glucose oxidase (GOD, EC: 1.1.3.4) entrapped in agarose-guar gum. The hydrophilic character of the agarose-guar gum composite matrix was checked by water contact angle measurement. The atomic force microscopy (AFM) images of the membranes showed proper confinement of both the enzymes during co-immobilization. The dynamic range for sucrose biosensor was achieved in the range of 1 x 10(-10) to 1 x 10(-7)M with lower detection limit 1 x 10(-10)M at pH 5.5 with 9 cycles of reuse. The spectrophotometric and electrochemical studies showed linear relationship between concentration of heavy metal ions and degree of inhibition of invertase. The toxicity sequence for invertase using both methods was observed as Hg(2+)>Pb(2+)>Ag(+)>Cd(2+). The dynamic linear range for mercury using electrochemical biosensor was observed in the range of 5 x 10(-10) to 12.5 x 10(-10)M for sucrose. The lower detection limit for the fabricated biosensor was found to be 5 x 10(-10)M. The reliability of the electrochemical biosensor was conformed by testing the spike samples and the results were comparable with the conventional photometric DNSA method. PMID:18667298

  19. Electrochemical biosensor for microRNA detection based on poly(U) polymerase mediated isothermal signal amplification.

    PubMed

    Zhou, Yunlei; Yin, Huanshun; Li, Jie; Li, Bingchen; Li, Xue; Ai, Shiyun; Zhang, Xiansheng

    2016-05-15

    MicroRNAs play crucial role in post-transcriptional regulation for gene expression in animals, plants, and viruses. For the better understanding of microRNA and its functions, it is very important to develop effectively analytical method for microRNA detection. Herein, a novel electrochemical biosensor was fabricated for sensitive and selective detection of microRNA based on poly(U) polymerase mediated isothermal signal amplification, where poly(U) polymerase can catalyze the template independent addition of UMP from UTP to the 3' end of RNA. Using this activity, the target microRNA can be successfully labeled with biotin conjugated UMPs at its 3'-end using biotin conjugated UTP (biotin-UTP) as donor. Then, the avidin conjugated alkaline phosphatase can be further captured to the 3'-end of the target microRNA based on the specific interaction between biotin and avidin. Finally, under the catalytic activity of alkaline phosphatase, the substrate of p-nitrophenyl phosphate disodium salt hexahydrate can be hydrolyzed to produce 4-nitrophenol. According to the relationship between the electrochemical signal of p-nitrophenol and the concentration of microRNA-319a, the content of microRNA-319a can be detected. This signal amplification method is simple and sensitive. The developed method can detect as low as 1.7 fM microRNA and produce precise and accurate linear dynamic range from 10 to 1000 fM. The fabricated biosensor was further applied to detect the expression level change of microRNA-319a in rice seedlings after incubation with five kinds of different phytohormones. PMID:26700579

  20. Alpha-Glucosidase Enzyme Biosensor for the Electrochemical Measurement of Antidiabetic Potential of Medicinal Plants.

    PubMed

    Mohiuddin, M; Arbain, D; Islam, A K M Shafiqul; Ahmad, M S; Ahmad, M N

    2016-12-01

    A biosensor for measuring the antidiabetic potential of medicinal plants was developed by covalent immobilization of α-glucosidase (AG) enzyme onto amine-functionalized multi-walled carbon nanotubes (MWCNTs-NH2). The immobilized enzyme was entrapped in freeze-thawed polyvinyl alcohol (PVA) together with p-nitrophenyl-α-D-glucopyranoside (PNPG) on the screen-printed carbon electrode at low pH to prevent the premature reaction between PNPG and AG enzyme. The enzymatic reaction within the biosensor is inhibited by bioactive compounds in the medicinal plant extracts. The capability of medicinal plants to inhibit the AG enzyme on the electrode correlates to the potential of the medicinal plants to inhibit the production of glucose from the carbohydrate in the human body. Thus, the inhibition indicates the antidiabetic potential of the medicinal plants. The performance of the biosensor was evaluated to measure the antidiabetic potential of three medicinal plants such as Tebengau (Ehretis laevis), Cemumar (Micromelum pubescens), and Kedondong (Spondias dulcis) and acarbose (commercial antidiabetic drug) via cyclic voltammetry, amperometry, and spectrophotometry. The cyclic voltammetry (CV) response for the inhibition of the AG enzyme activity by Tebengau plant extracts showed a linear relation in the range from 0.423-8.29 μA, and the inhibition detection limit was 0.253 μA. The biosensor exhibited good sensitivity (0.422 μA/mg Tebengau plant extracts) and rapid response (22 s). The biosensor retains approximately 82.16 % of its initial activity even after 30 days of storage at 4 °C. PMID:26887579

  1. Alpha-Glucosidase Enzyme Biosensor for the Electrochemical Measurement of Antidiabetic Potential of Medicinal Plants

    NASA Astrophysics Data System (ADS)

    Mohiuddin, M.; Arbain, D.; Islam, A. K. M. Shafiqul; Ahmad, M. S.; Ahmad, M. N.

    2016-02-01

    A biosensor for measuring the antidiabetic potential of medicinal plants was developed by covalent immobilization of α-glucosidase (AG) enzyme onto amine-functionalized multi-walled carbon nanotubes (MWCNTs-NH2). The immobilized enzyme was entrapped in freeze-thawed polyvinyl alcohol (PVA) together with p-nitrophenyl-α- d-glucopyranoside (PNPG) on the screen-printed carbon electrode at low pH to prevent the premature reaction between PNPG and AG enzyme. The enzymatic reaction within the biosensor is inhibited by bioactive compounds in the medicinal plant extracts. The capability of medicinal plants to inhibit the AG enzyme on the electrode correlates to the potential of the medicinal plants to inhibit the production of glucose from the carbohydrate in the human body. Thus, the inhibition indicates the antidiabetic potential of the medicinal plants. The performance of the biosensor was evaluated to measure the antidiabetic potential of three medicinal plants such as Tebengau ( Ehretis laevis), Cemumar ( Micromelum pubescens), and Kedondong ( Spondias dulcis) and acarbose (commercial antidiabetic drug) via cyclic voltammetry, amperometry, and spectrophotometry. The cyclic voltammetry (CV) response for the inhibition of the AG enzyme activity by Tebengau plant extracts showed a linear relation in the range from 0.423-8.29 μA, and the inhibition detection limit was 0.253 μA. The biosensor exhibited good sensitivity (0.422 μA/mg Tebengau plant extracts) and rapid response (22 s). The biosensor retains approximately 82.16 % of its initial activity even after 30 days of storage at 4 °C.

  2. Triple-helix DNA structural studies using a Love wave acoustic biosensor.

    PubMed

    Papadakis, George; Tsortos, Achilleas; Gizeli, Electra

    2009-12-15

    The development of sensors for detecting the conformation of surface-attached molecules is an emerging field with significance in the pharmaceutical industry and in drug design. In this work, triplex-forming oligos (TFOs), a separate class of non-natural DNA bending agents that can affect the mechanical properties of DNA through the formation of triple-helical structures of specific conformation and/or flexibility, are used as a model system in combination with an acoustic biosensor to determine molecular geometrical features. In practice, the degree of bending of a specific DNA target caused by a particular TFO was evaluated by measuring the ratio of acoustic energy change over phase change observed during the binding of pre-formed triplex DNA molecules to the device surface. The DNA bending angle derived via acoustic measurements is in excellent agreement with previously reported values using molecular biology techniques. The reported acoustic technique appears quite appealing for the biophysical study of DNA molecules providing rapid qualitative and quantitative information, at the same time holding promise to be developed as a high-throughput method for the evaluation of DNA conformational changes. PMID:19748772

  3. Development of an Automated DNA Detection System Using an Electrochemical DNA Chip Technology

    NASA Astrophysics Data System (ADS)

    Hongo, Sadato; Okada, Jun; Hashimoto, Koji; Tsuji, Koichi; Nikaido, Masaru; Gemma, Nobuhiro

    A new compact automated DNA detection system Genelyzer™ has been developed. After injecting a sample solution into a cassette with a built-in electrochemical DNA chip, processes from hybridization reaction to detection and analysis are all operated fully automatically. In order to detect a sample DNA, electrical currents from electrodes due to an oxidization reaction of electrochemically active intercalator molecules bound to hybridized DNAs are detected. The intercalator is supplied as a reagent solution by a fluid supply unit of the system. The feasibility test proved that the simultaneous typing of six single nucleotide polymorphisms (SNPs) associated with a rheumatoid arthritis (RA) was carried out within two hours and that all the results were consistent with those by conventional typing methods. It is expected that this system opens a new way to a DNA testing such as a test for infectious diseases, a personalized medicine, a food inspection, a forensic application and any other applications.

  4. Nanoparticle-based DNA biosensor for visual detection of genetically modified organisms.

    PubMed

    Kalogianni, Despina P; Koraki, Theodora; Christopoulos, Theodore K; Ioannou, Penelope C

    2006-01-15

    Although screening of raw ingredients and food products for genetically modified organisms (GMO) may be accomplished by detecting either the exogenous DNA or the novel protein, DNA is the preferred analyte because of its superior stability during food processing. The development of DNA biosensors is of increasing importance due to the growing demand for rapid and reliable methods for GMO detection. We report the first DNA biosensor in a dry-reagent dipstick configuration for visual detection and confirmation of GMO-related sequences by hybridization within minutes. The sensor is disposable and does not require special instrumentation. It detects the 35S promoter and nopaline synthase (NOS) terminator sequences that are present in the majority of transgenic plants. The target sequences are amplified by the polymerase chain reaction (PCR) and hybridized (7min) with probes bearing oligo(dA) tail. The biotinylated product is applied to the sensor followed by immersion in the appropriate buffer. Migration of the buffer rehydrates gold nanoparticles conjugated to oligo(dT), which hybridize with the oligo(dA) tails. The hybrids are captured by immobilized streptavidin at the test zone of the sensor giving a characteristic red line due to the accumulation of the nanoparticles. The excess of nanoparticle conjugates are captured at the control zone by immobilized oligo(dA) strands. Amplified 35S or NOS DNA is detectable at 0.16nM. Soybean powder certified reference material with 0.1% GMO content is clearly detectable after 35 and 40 amplification cycles for 35S and NOS sequence, respectively. The sensor was also applied to real samples from various sources. PMID:15935636

  5. Mechanism for invalid detection of microcantilever-DNA biosensors due to environmental changes

    NASA Astrophysics Data System (ADS)

    Tan, Z.-Q.; Zhang, N.-H.; Meng, W.-L.; Tang, H.-S.

    2016-06-01

    Microcantilever-DNA biosensors can lose recognition signals under specific hybridization conditions; this could be termed as a type of invalid detection. Using a multiscale energy method, this paper presents an alternative mechanism for this invalid detection induced by bio-interactions and environmental changes in temperature and ionic strength. First, a scaling law for the nanoscale thickness of the DNA film, and a mesoscopic empirical potential for bio-interactions in DNA liquid crystal solution, were combined to update a multiscale analytical model revealing the relation between cantilever motion, temperature, ionic strength, elastic properties of multilayered cantilevers, and nanoscopic properties of DNA molecules. Second, we carried out isothermal and non-isothermal experiments for the bending motion during the formation of a self-assembled monolayer of thiolated single-stranded DNA covalently immobilized on the gold-coated side of the cantilevers, and during the subsequent hybridization with the complementary nucleic acid, in order to obtain the relevant model parameters, and also to validate the proposed analytical model. Third, the effects of temperature and ionic strength on the microcantilever deflections were investigated. Numerical results show that the competing interplay among electrostatic force, hydration force, and configurational entropy generates an invalid point of detection at a grafting density of about 0.05 chain nm‑2. In the grafting density interval of 0.02–0.05 chain nm‑2, the thermal effect induces distortion of signals; in the grafting density interval of 0.05–0.097 chain nm‑2, fluctuations in ionic strength make detection fail. These findings will help to design and improve microcantilever-based biosensors with high sensitivity and robustness.

  6. New Catalytic DNA Biosensors for Radionuclides and Metal ions

    SciTech Connect

    Lu, Yi

    2005-06-01

    In vitro selection for DNAzymes that are catalytically active with UO22+ ions as the metal cofactor has been completed. The 10th generation pool of DNA was cloned and sequenced. A total of 84 clones were sequenced and placed into families based on sequence alignments. Selected members of each family were 5-labeled with 32P and amplified using PCR. Activity assays were conducted using the isotopically labeled DNAzymes in order to determine which sequences were the most active. The secondary structures of the two most active sequences, called Clone 13 and Clone 39, were determined using the computer program Mfold. A cleavage rate of approximately 1 min-1 in the presence of 10 uM UO22+ was observed for both clones. Clone 39 was determined to be the best candidate for truncation to create a trans-cleaving DNAzyme, based on its secondary structure. An enzyme strand, called 39E, and a substrate strand, called 39DS, were designed by truncating the cis-cleaving DNAzyme. An alternative enzyme strand, called 39Ec, was also assayed with the 39DS substrate. This strand was designed so that the two binding arms were perfectly complimentary, unlike 39E, which formed three mismatched base pairs with 39DS. Both 39E and 39Ec were found to be active, with a rate of approximately 1 min-1 in the presence of 10 uM UO22+. A preliminary UO22+ binding curve was obtained for the 39Ec/39DS trans-cleaving system. The enzyme is active with UO22+ concentrations as low as 1 nM. Based on the preliminary binding curve data, the apparent UO22+ binding constant is approximately 330 nM, and kmax is approximately 1 min-1.

  7. Highly-sensitive liquid crystal biosensor based on DNA dendrimers-mediated optical reorientation.

    PubMed

    Tan, Hui; Li, Xia; Liao, Shuzhen; Yu, Ruqin; Wu, Zhaoyang

    2014-12-15

    A novel highly-sensitive liquid crystal (LC) biosensing approach based on target-triggering DNA dendrimers was developed for the detection of p53 mutation gene segment at the LC-aqueous interface. In this study, the mutant-type p53 gene segment was the target to trigger the formation of DNA dendrimers from hairpin DNA probes by hybridization chain reaction, and the latter as a 'signal enhancement element' further induced the LC reorientation from tilted to homeotropic alignment, resulting in a corresponding optical changes of LC biosensors from birefringent to honeycombed textures or dark framework. The distinct optical reorientational appearances can serve as a characteristic signal to distinguish target concentrations ranging from 0.08 nM to 8 nM. Moreover, these optical phenomena suggest that the LC reorientation is related to the electric-dipole coupling between the adsorbed DNA and LC molecules, the conformational constraints of DNA and the internal electric field induction upon hybridization. This label-free LC biosensing strategy can open up a new platform for the sensitive detection of specific DNA sequences and enrich the application scope of an LC biosensing technique. PMID:24984288

  8. REVIEW ARTICLE: Environmental applications of analytical biosensors

    NASA Astrophysics Data System (ADS)

    Marco, María-Pilar; Barceló, Damià

    1996-11-01

    A review of the fundamental aspects and environmental applications of biosensors is presented. The bases of different transducer principles such as electrochemical, optical and piezoelectric are discussed. Various examples are given of the applications of such principles to develop immunosensor devices to determine common environmental contaminants. Attention is also paid to catalytic biosensors, using enzymes as sensing elements. Biosensor devices based on the use of cholinesterase and various oxidase enzymes such as tyrosinase, laccase, peroxidase and aldehyde dehydrogenase are reported. Some examples are given of the applications of other biomolecules such as whole cells, DNA or proteins, to determine pollution. Validation studies are presented comparing biosensors with chromatographic techniques to determine organophosphorus pesticides and phenolic compounds in environmental samples.

  9. Label-free electrochemical impedance biosensor to detect human interleukin-8 in serum with sub-pg/ml sensitivity

    PubMed Central

    Sharma, R.; Deacon, S.E.; Nowak, D.; George, S.E.; Szymonik, M.P.; Tang, A.A.S.; Tomlinson, D.C.; Davies, A.G.; McPherson, M.J.; Wälti, C.

    2016-01-01

    Biosensors with high sensitivity and short time-to-result that are capable of detecting biomarkers in body fluids such as serum are an important prerequisite for early diagnostics in modern healthcare provision. Here, we report the development of an electrochemical impedance-based sensor for the detection in serum of human interleukin-8 (IL-8), a pro-angiogenic chemokine implicated in a wide range of inflammatory diseases. The sensor employs a small and robust synthetic non-antibody capture protein based on a cystatin scaffold that displays high affinity for human IL-8 with a KD of 35±10 nM and excellent ligand specificity. The change in the phase of the electrochemical impedance from the serum baseline, ∆θ(ƒ), measured at 0.1 Hz, was used as the measure for quantifying IL-8 concentration in the fluid. Optimal sensor signal was observed after 15 min incubation, and the sensor exhibited a linear response versus logarithm of IL-8 concentration from 900 fg/ml to 900 ng/ml. A detection limit of around 90 fg/ml, which is significantly lower than the basal clinical levels of 5–10 pg/ml, was observed. Our results are significant for the development of point-of-care and early diagnostics where high sensitivity and short time-to-results are essential. PMID:26897263

  10. CdS quantum dots modified CuO inverse opal electrodes for ultrasensitive electrochemical and photoelectrochemical biosensor

    PubMed Central

    Xia, Lei; Xu, Lin; Song, Jian; Xu, Ru; Liu, Dali; Dong, Biao; Song, Hongwei

    2015-01-01

    The CuO inverse opal photonic crystals (IOPCs) were synthesized by the sol-gel method and modified with CdS quantum dots by successive ionic layer adsorption and reaction (SILAR). CdS QDs modified CuO IOPCs FTO electrodes of different SILAR cycles were fabricated and their electrochemical properties were studied by cyclic voltammetry (CV) and chronoamperometry (I–t). Structure and morphology of the samples were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), high-resolution TEM (HRTEM), Energy-dispersive X-ray analysis (EDX) and X-ray diffraction pattern (XRD). The result indicated that the structure of IOPCs and loading of CdS QDs could greatly improve the electrochemical properties. Three SILAR cycles of CdS QDs sensitization was the optimum condition for preparing electrodes, it exhibited a sensitivity of 4345 μA mM-1 cm-2 to glucose with a 0.15 μM detection limit (S/N= 3) and a linear range from 0.15 μM to 0.5 mM under a working potential of +0.7 V. It also showed strong stability, good reproducibility, excellent selectivity and fast amperometric response. This work provides a promising approach for realizing excellent photoelectrochemical nonenzymatic glucose biosensor of similar composite structure. PMID:26042520

  11. Electrochemical functionalization of gold and silicon surfaces by a maleimide group as a biosensor for immunological application.

    PubMed

    Zhang, Xin; Tretjakov, Aleksei; Hovestaedt, Marc; Sun, Guoguang; Syritski, Vitali; Reut, Jekaterina; Volkmer, Rudolf; Hinrichs, Karsten; Rappich, Joerg

    2013-03-01

    In the present study we investigated the preparation of biofunctionalized surfaces using the direct electrochemical grafting of maleimidophenyl molecules with subsequent covalent immobilization of specific peptide to detect target antibody, thereby extending the application of the biosensing systems towards immunodiagnostics. Para-maleimidophenyl (p-MP) functional groups were electrochemically grafted on gold and silicon surfaces from solutions of the corresponding diazonium salt. A specially synthesized peptide modified with cysteine (Cys-peptide) was then immobilized on the p-MP grafted substrates by cross-linking between the maleimide groups and the sulfhydryl group of the cysteine residues. Accordingly, the Cys-peptide worked as an antigen that was able to bind specifically the target antibody (anti-GST antibody), while it was non-sensitive to a negative contrast antibody (i.e. anti-Flag β). The immobilization of both specific and non-specific antibodies on the Cys-peptide-modified surfaces was monitored by infrared spectroscopic ellipsometry, a quartz crystal microbalance integrated in flow injection analysis system and potentiometric response. The results obtained clearly demonstrated that the direct modification of a surface with maleimidophenyl provides a very simple and reliable way of preparing biofunctionalized surfaces suitable for the construction of immunological biosensors. PMID:23117146

  12. Label-free electrochemical impedance biosensor to detect human interleukin-8 in serum with sub-pg/ml sensitivity.

    PubMed

    Sharma, R; Deacon, S E; Nowak, D; George, S E; Szymonik, M P; Tang, A A S; Tomlinson, D C; Davies, A G; McPherson, M J; Wälti, C

    2016-06-15

    Biosensors with high sensitivity and short time-to-result that are capable of detecting biomarkers in body fluids such as serum are an important prerequisite for early diagnostics in modern healthcare provision. Here, we report the development of an electrochemical impedance-based sensor for the detection in serum of human interleukin-8 (IL-8), a pro-angiogenic chemokine implicated in a wide range of inflammatory diseases. The sensor employs a small and robust synthetic non-antibody capture protein based on a cystatin scaffold that displays high affinity for human IL-8 with a KD of 35 ± 10 nM and excellent ligand specificity. The change in the phase of the electrochemical impedance from the serum baseline, ∆θ(ƒ), measured at 0.1 Hz, was used as the measure for quantifying IL-8 concentration in the fluid. Optimal sensor signal was observed after 15 min incubation, and the sensor exhibited a linear response versus logarithm of IL-8 concentration from 900 fg/ml to 900 ng/ml. A detection limit of around 90 fg/ml, which is significantly lower than the basal clinical levels of 5-10 pg/ml, was observed. Our results are significant for the development of point-of-care and early diagnostics where high sensitivity and short time-to-results are essential. PMID:26897263

  13. CdS quantum dots modified CuO inverse opal electrodes for ultrasensitive electrochemical and photoelectrochemical biosensor.

    PubMed

    Xia, Lei; Xu, Lin; Song, Jian; Xu, Ru; Liu, Dali; Dong, Biao; Song, Hongwei

    2015-01-01

    The CuO inverse opal photonic crystals (IOPCs) were synthesized by the sol-gel method and modified with CdS quantum dots by successive ionic layer adsorption and reaction (SILAR). CdS QDs modified CuO IOPCs FTO electrodes of different SILAR cycles were fabricated and their electrochemical properties were studied by cyclic voltammetry (CV) and chronoamperometry (I-t). Structure and morphology of the samples were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), high-resolution TEM (HRTEM), Energy-dispersive X-ray analysis (EDX) and X-ray diffraction pattern (XRD). The result indicated that the structure of IOPCs and loading of CdS QDs could greatly improve the electrochemical properties. Three SILAR cycles of CdS QDs sensitization was the optimum condition for preparing electrodes, it exhibited a sensitivity of 4345 μA mM(-1) cm(-2) to glucose with a 0.15 μM detection limit (S/N= 3) and a linear range from 0.15 μM to 0.5 mM under a working potential of +0.7 V. It also showed strong stability, good reproducibility, excellent selectivity and fast amperometric response. This work provides a promising approach for realizing excellent photoelectrochemical nonenzymatic glucose biosensor of similar composite structure. PMID:26042520

  14. Preparation of novel mercury-doped silver nanoparticles film glassy carbon electrode and its application for electrochemical biosensor.

    PubMed

    Li, Mao-Guo; Shang, Yong-Jia; Gao, Ying-Chun; Wang, Guang-Feng; Fang, Bin

    2005-06-01

    A novel mercury-doped silver nanoparticles film glassy carbon (Ag/MFGC) electrode was prepared in this study. Electrochemical behaviors of cysteine on the Ag/MFGC electrode were investigated by electrochemical impedance spectroscopy and cyclic voltammetry (CV). The results indicated that cysteine could be strongly adsorbed on the surface of the Ag/MFGC electrode to form a thin layer. The doped electrode could catalyze the electrode reaction process of cysteine, and the cysteine displayed a pair of well-defined and nearly reversible CV peaks at the electrode in an acetate buffer solution (pH 5.0). The Ag/MFGC electrode was used for determination of cysteine by differential pulse voltammetry. The linear range was between 4.0x10(-7) and 1.3x10(-5) mol/L, with a detection limit of 1.0x10(-7) mol/L and a signal-to-noise ratio of 3. The relative standard deviation was 2.4% for seven successive determinations of 1.0x10(-5) mol/L cysteine. The determinations of cysteine in synthetic samples and urinal samples were carried out and satisfactory results were obtained. Amperometric application of the Ag/MFGC electrode as biosensors is proposed. PMID:15866527

  15. A highly sensitive electrochemical biosensor based on zinc oxide nanotetrapods for L-lactic acid detection.

    PubMed

    Lei, Yang; Luo, Ning; Yan, Xiaoqin; Zhao, Yanguang; Zhang, Gong; Zhang, Yue

    2012-06-01

    An amperometric biosensor based on zinc oxide (ZnO) nanotetrapods was designed to detect L-lactic acid. The lactate oxidase was immobilized on the surface of ZnO nanotetrapods by electrostatic adsorption. Unlike traditional detectors, the special four-leg individual ZnO nanostructure, as an adsorption layer, provides multiterminal charge transfer channels. Furthermore, a large amount of ZnO tetrapods are randomly stacked to form a three-dimensional network naturally that facilitates the exchange of electrons and ions in the phosphate buffer solution. Utilizing amperometric response measurements, the prepared ZnO nanotetrapod L-lactic acid biosensor displayed a detection limit of 1.2 μM, a low apparent Michaelis-Menten constant of 0.58 mM, a high sensitivity of 28.0 μA cm(-2) mM(-1) and a good linear relationship in the range of 3.6 μM-0.6 mM for the L-lactic acid detection. This study shows that the biosensor based on ZnO tetrapod nanostructures is highly sensitive and able to respond rapidly in detecting lactic acid. PMID:22538963

  16. A highly oriented hybrid microarray modified electrode fabricated by a template-free method for ultrasensitive electrochemical DNA recognition.

    PubMed

    Shi, Lei; Chu, Zhenyu; Dong, Xueliang; Jin, Wanqin; Dempsey, Eithne

    2013-11-01

    Highly oriented growth of a hybrid microarray was realized by a facile template-free method on gold substrates for the first time. The proposed formation mechanism involves an interfacial structure-directing force arising from self-assembled monolayers (SAMs) between gold substrates and hybrid crystals. Different SAMs and variable surface coverage of the assembled molecules play a critical role in the interfacial directing forces and influence the morphologies of hybrid films. A highly oriented hybrid microarray was formed on the highly aligned and vertical SAMs of 1,4-benzenedithiol molecules with rigid backbones, which afforded an intense structure-directing power for the oriented growth of hybrid crystals. Additionally, the density of the microarray could be adjusted by controlling the surface coverage of assembled molecules. Based on the hybrid microarray modified electrode with a large specific area (ca. 10 times its geometrical area), a label-free electrochemical DNA biosensor was constructed for the detection of an oligonucleotide fragment of the avian flu virus H5N1. The DNA biosensor displayed a significantly low detection limit of 5 pM (S/N = 3), a wide linear response from 10 pM to 10 nM, as well as excellent selectivity, good regeneration and high stability. We expect that the proposed template-free method can provide a new reference for the fabrication of a highly oriented hybrid array and the as-prepared microarray modified electrode will be a promising paradigm in constructing highly sensitive and selective biosensors. PMID:24061929

  17. Non-enzymatic electrochemical biosensor based on Pt NPs/RGO-CS-Fc nano-hybrids for the detection of hydrogen peroxide in living cells.

    PubMed

    Bai, Zhihao; Li, Guiyin; Liang, Jingtao; Su, Jing; Zhang, Yue; Chen, Huaizhou; Huang, Yong; Sui, Weiguo; Zhao, Yongxiang

    2016-08-15

    A highly sensitive non-enzymatic electrochemical sensor based on platinum nanoparticles/reduced graphene oxide-chitosan-ferrocene carboxylic acid nano-hybrids (Pt NPs/RGO-CS-Fc biosensor) was developed for the measurement of hydrogen peroxide (H2O2). The RGO-CS-Fc nano-hybrids was prepared and characterized by UV-vis spectrum, Fourier transform infrared spectroscopy, transmission electron microscopy, Raman spectrometer and electrochemical impedance spectroscopy. Under optimal experimental conditions, the Pt NPs/RGO-CS-Fc biosensor showed outstanding catalytic activity toward H2O2 reduction. The current response of the biosensor presented a linear relationship with H2O2 concentration from 2.0×10(-8)M to 3.0×10(-6)M with a correlation coefficient of R(2)=0.9968 and with logarithm of H2O2 concentration from 6.0×10(-6)M to 1.0×10(-2)M with a correlation coefficient of R(2)=0.9887, the low detection limit of 20nM was obtained at the signal/noise (S/N) ratio of 3. Moreover, the Pt NPs/RGO-CS-Fc biosensor exhibited excellent anti-interference capability and reproducibility for the detection of H2O2. The biosensor was also successfully applied for the detection of H2O2 from living cells containing normal and cancer cells. All these results prove that the Pt NPs/RGO-CS-Fc biosensor has the potential application in clinical diagnostics to evaluate oxidative stress of different living cells. PMID:27085950

  18. New highly sensitive and selective catalytic DNA biosensors for metal ions.

    PubMed

    Lu, Yi; Liu, Juewen; Li, Jing; Bruesehoff, Peter J; Pavot, Caroline M-B; Brown, Andrea K

    2003-05-01

    While remarkable progress has been made in developing sensors for metal ions such as Ca(II) and Zn(II), designing and synthesizing sensitive and selective metal ion sensors remains a significant challenge. Perhaps the biggest challenge is the design and synthesis of a sensor capable of specific and strong metal binding. Since our knowledge about the construction of metal-binding sites in general is limited, searching for sensors in a combinatorial way is of significant value. Therefore, we have been able to use a combinatorial method called in vitro selection to obtain catalytic DNA that can bind a metal ion of choice strongly and specifically. The metal ion selectivity of the catalytic DNA was further improved using a 'negative selection' strategy where catalytic DNA that are selective for competing metal ions are discarded in the in vitro selection processes. By labeling the resulting catalytic DNA with a fluorophore/quencher pair, we have made a new class of metal ion fluorescent sensors that are the first examples of catalytic DNA biosensors for metal ions. The sensors combine the high selectivity of catalytic DNA with the high sensitivity of fluorescent detection, and can be applied to the quantitative detection of metal ions over a wide concentration range and with high selectivity. The use of DNA sensors in detection and quantification of lead ions in environmental samples such as water from Lake Michigan has been demonstrated. DNA is stable, cost-effective, environmentally benign, and easily adaptable to optical fiber and microarray technology for device manufacture. Thus, the DNA sensors explained here hold great promise for on-site and real-time monitoring of metal ions in the fields of environmental monitoring, developmental biology, clinical toxicology, wastewater treatment, and industrial process monitoring. PMID:12706559

  19. Electrochemical Sensors for Clinic Analysis

    PubMed Central

    Wang, You; Xu, Hui; Zhang, Jianming; Li, Guang

    2008-01-01

    Demanded by modern medical diagnosis, advances in microfabrication technology have led to the development of fast, sensitive and selective electrochemical sensors for clinic analysis. This review addresses the principles behind electrochemical sensor design and fabrication, and introduces recent progress in the application of electrochemical sensors to analysis of clinical chemicals such as blood gases, electrolytes, metabolites, DNA and antibodies, including basic and applied research. Miniaturized commercial electrochemical biosensors will form the basis of inexpensive and easy to use devices for acquiring chemical information to bring sophisticated analytical capabilities to the non-specialist and general public alike in the future.

  20. A novel label-free and enzyme-free electrochemical aptasensor based on DNA in situ metallization.

    PubMed

    Qian, Yong; Gao, Fenglei; Du, Lili; Zhang, Yu; Tang, Daoquan; Yang, Dongzhi

    2015-12-15

    In this work, we presented a novel label-free and enzyme-free electrochemical aptasensor based on DNA in situ silver metallization as effective electrochemical label. Molecular beacon 2 (MB2, Peptide nucleic acid) was first immobilized on the gold electrode (AuE) through Au-S bond. In the presence of thrombin, the thrombin binding aptamer (MB1) preferred to form thrombin/aptamer complex in lieu of aptamer-DNA duplex, resulting in the 8-17 DNAzyme liberating from the caged structure and hybridization with the MB2, the MB2 will replace and free the target thrombin when it hybridizes with MB1. The released target thrombin can participate in the next hybridization process with MB1. Eventually, each target thrombin went through many cycles, resulting in numerous MB1 confining close to the AuE, which leaded to the surface became negatively charged and allowed the absorption of silver ions on the DNA skeleton. After chemical reduction by hydroquinone, the formed silver nanoparticles could be afforded a signal trace for electrochemical stripping analysis of target thrombin. Through introducing a hybridization chain reaction to increase the DNA length, the current signal was further amplified, achieved the detection of thrombin with a linear range from 1.0×10(-16) to 1.0×10(-11) M and a detection limit of 37 aM. In addition, the signal amplification is realized without using any enzymes or sophisticated label process, and the sensing strategy is completely non-labeled. The success in the present biosensor served as a significant step towards the development of monitoring ultratrace thrombin in clinical detection. PMID:26176208

  1. A single electrochemical biosensor for detecting the activity and inhibition of both protein kinase and alkaline phosphatase based on phosphate ions induced deposition of redox precipitates.

    PubMed

    Shen, Congcong; Li, Xiangzhi; Rasooly, Avraham; Guo, Linyan; Zhang, Kaina; Yang, Minghui

    2016-11-15

    Protein kinase (PKA) and alkaline phosphatase (ALP) are clinically relevant enzymes for a number of diseases. In this work, we developed a new simple electrochemical biosensor for the detection of the activity and inhibition of both PKA and ALP. One common feature of the PKA and ALP catalyzing process is that PKA can hydrolysis adenosine-5'-triphosphate (ATP) and ALP can hydrolysis pyrophosphate, both reactions produce phosphate ions, and the amount of phosphate ion produced is proportional to enzyme activity. Our assay is based on the principle that phosphate ions react with molybdate to form redox molybdophosphate precipitates on the electrode surface, thus generating electrochemical current. The detection limit for PKA and ALP were much lower than existing assays. The biosensor has good specificity and was used to measure drug-stimulated PKA from lysates of HeLa cells. We also evaluated the use of the biosensor as a screening tool for enzyme inhibitors. To the best of our knowledge, this is the first report of a biosensor capable of detecting the activity of both PKA and ALP. This tool has the potential to simplify PKA and ALP clinical measurement, thereby improving diagnostics of relevant diseases. It also may serve as the basis for a simple screening method for new enzyme inhibitors for disease treatment. PMID:27179562

  2. Ultrasensitive electrochemical biosensor based on reduced graphene oxide-tetraethylene pentamine-BMIMPF6 hybrids for the detection of α2,6-sialylated glycans in human serum.

    PubMed

    Li, Yuliang; He, Junlin; Niu, Yazhen; Yu, Chao

    2015-12-15

    In this paper, a simple, ultrasensitive and label-free electrochemical α2,6-sialylated glycans biosensor based on reduced graphene oxide-tetraethylene pentamine-1-butyl-3-methylimidazolium hexafluorophosphate (BMIMPF6) hybrids was developed. Due to the abundance of amino groups from reduced graphene oxide-tetraethylene pentamine (rGO-TEPA) and the electrostatic interaction of BMIMPF6, bimetallic gold platinum alloy nanoparticles (AuPtNPs) were densely adsorbed onto the surface of the nanocomposite, providing a large surface area available for the immobilization of Sambucus nigra agglutinin (SNA). AuPtNPs have excellent conductivity and catalytic activity, which can promote electron transfer between the electrolyte solution and the surface of electrode and can enhance the sensitivity of biosensor. SNA, which specifically binds α2,6-sialylated glycans, was covalently immobilized on AuPtNPs for specific detection of α2,6-sialylated glycans in human serum. Under optimal experimental conditions, amperometric response changes were used to detect α2,6-sialylated glycans with a broad linear range of 10 fg mL(-1) -1 μg mL(-1) and a low detection limit of 3 fg mL(-1) (S/N=3). When applied to spiked serum samples, the recovery of the developed biosensor ranged from 100.8% to 101.4%, suggesting that the electrochemical biosensor would be suitable for the practical detection of α2,6-sialylated glycans. PMID:26257188

  3. DNA-functionalized upconversion nanoparticles as biosensors for rapid, sensitive, and selective detection of Hg(2+) in complex matrices.

    PubMed

    Huang, Li-Jiao; Yu, Ru-Qin; Chu, Xia

    2015-08-01

    We have developed a facile one-step approach to make hydrophilic and DNA-functionalizable upconversion nanoparticles (UCNPs), which are used to act as a biosensor for determining Hg(2+) in complex matrices. The proposed approach is simple and exhibits low background interference, high sensitivity and rapid response. PMID:26040459

  4. A Sensitive and simple macrophage-based electrochemical biosensor for evaluating lipopolysaccharide cytotoxicity of pathogenic bacteria.

    PubMed

    Wang, Xiumei; Zhu, Pei; Pi, Fuwei; Jiang, Hui; Shao, Jingdong; Zhang, Yinzhi; Sun, Xiulan

    2016-07-15

    In this study, a sensitive and simple electrochemical murine macrophage (Ana-1) cell sensor has been developed for early detection of lipopolysaccharides (LPS) to evaluate the toxicity of pathogenic bacteria. Magnetic glassy carbon electrode (MGCE), which possesses excellent reproducibility and regeneration qualities, was modified with a nanocomposite to improve electrochemical signals and enhance the sensitivity. The synthesized magnetic nanoparticles (MNPs) were internalized into murine macrophages, which completed the immobilization of macrophages onto the modified electrode for evaluating the cytotoxicity of LPS by electrochemical impedance spectroscopy (EIS). The MNPs facilitated reusability of the proposed sensor by allowing removal of the magnetic core from the electrode. Our results indicated that LPS caused a marked decrease in electrochemical impedance in a dose-dependent manner in range of 1-5μg/mL. By SEM, we found that microvilli on the plasma membrane became scarce and the membrane became smooth on cells incubated with LPS, which lessens the absorption of cells to reduce the impedance. And biological assay indicated that EIS patterns were correlated with the calcium concentration in cells, and suggested that [Ca(2+)]i production increased in cells incubated with LPS and its mobilization altered electrochemical signals. Compared with conventional methods, this electrochemical test is inexpensive, highly sensitive, and has a quick response, and thus provides a new avenue for evaluating the cytotoxicity of pathogens. PMID:26991601

  5. Electrochemical signature of mismatch in overhang DNA films: a scanning electrochemical microscopic study.

    PubMed

    Shamsi, Mohtashim Hassan; Kraatz, Heinz-Bernhard

    2013-06-21

    High throughput DNA basepair mismatch detection is an ultimate goal for earlier and point-of-care diagnostics. However, the size of a target sequence on single nucleotide mismatch detection will critically impact the design of sensors in future. To study the potential impact of target size, the probe and target strands of unequal size were hybridized in the absence and presence of single nucleotide mismatches along the sequence. After hybridization, the shorter target sequences form overhangs in the probe strand while longer target sequences form overhangs in the complementary strand. The resulting double stranded DNA hybrids were printed on gold surfaces and the electrochemical response of the films was studied by scanning electrochemical microscopy without signal amplification and label. The redox mediator, [Fe(CN)(6)](4-), experiences lower repulsion in the vicinity of mismatch containing ds-DNA films, which ultimately manifests into higher feedback current regardless of the size and hybridization position of the complementary strands. Kinetic rate constants monitored right above the ds-DNA films show k(0) = 4.5 ± 0.1 × 10(-5) cm s(-1) for the short sequence hybridized at the upper portion of the probe while k(0) = 4.1 ± 0.2 × 10(-5) cm s(-1) for longer complementary strands which has only top overhang. It suggests that hybridization position is important for mismatch detection in short complementary stands. However, in longer complementary strands, mismatches are easily detectable in the absence of bottom overhangs. PMID:23671908

  6. Ultrasensitive electrochemical detection of avian influenza A (H7N9) virus DNA based on isothermal exponential amplification coupled with hybridization chain reaction of DNAzyme nanowires.

    PubMed

    Yu, Yanyan; Chen, Zuanguang; Jian, Wensi; Sun, Duanping; Zhang, Beibei; Li, Xinchun; Yao, Meicun

    2015-02-15

    In this work, a simple and label-free electrochemical biosensor with duel amplification strategy was developed for DNA detection based on isothermal exponential amplification (EXPAR) coupled with hybridization chain reaction (HCR) of DNAzymes nanowires. Through rational design, neither the primer nor the DNAzymes containing molecular beacons (MBs) could react with the duplex probe which were fixed on the electrode surface. Once challenged with target, the duplex probe cleaved and triggered the EXPAR mediated target recycle and regeneration circles as well as the HCR process. As a result, a greater amount of targets were generated to cleave the duplex probes. Subsequently, the nanowires consisting of the G-quadruplex units were self-assembled through hybridization with the strand fixed on the electrode surface. In the presence of hemin, the resulting catalytic G-quadruplex-hemin HRP-mimicking DNAzymes were formed. Electrochemical signals can be obtained by measuring the increase in reduction current of oxidized 3.3',5.5'-tetramethylbenzidine sulfate (TMB), which was generated by DNAzyme in the presence of H2O2. This method exhibited ultrahigh sensitivity towards avian influenza A (H7N9) virus DNA sequence with detection limits of 9.4 fM and a detection range of 4 orders of magnitude. The biosensor was also capable of discriminating single-nucleotide difference among concomitant DNA sequences and performed well in spiked cell lysates. PMID:25310490

  7. A signal-on electrochemical aptasensor for ultrasensitive detection of endotoxin using three-way DNA junction-aided enzymatic recycling and graphene nanohybrid for amplification

    NASA Astrophysics Data System (ADS)

    Bai, Lijuan; Chai, Yaqin; Pu, Xiaoyun; Yuan, Ruo

    2014-02-01

    Endotoxin, also known as lipopolysaccharide (LPS), is able to induce a strong immune response on its internalization into mammalian cells. To date, aptamer-based biosensors for LPS detection have been rarely reported. This work describes a new signal-on electrochemical aptasensor for the ultrasensitive detection of LPS by combining the three-way DNA hybridization process and nanotechnology-based amplification. With the help of DNA1 (associated with the concentration of target LPS), the capture probe hybridizes with DNA1 and the assistant probe to open its hairpin structure and form a ternary ``Y'' junction structure. The DNA1 can be released from the structure in the presence of nicking endonuclease to initiate the next hybridization process. Then a great deal of cleaved capture probe produced in the cyclic process can bind with DNA2-nanocomposite, which contains the electroactive toluidine blue (Tb) with the amplification materials graphene (Gra) and gold nanoparticles (AuNPs). Thus, an enhanced electrochemical signal can be easily read out. With the cascade signal amplification, this newly designed protocol provides an ultrasensitive electrochemical detection of LPS down to the femtogram level (8.7 fg mL-1) with a linear range of 6 orders of magnitude (from 10 fg mL-1 to 50 ng mL-1). Moreover, the high sensitivity and specificity make this method versatile for the detection of other biomolecules by changing the corresponding sequences of the capture probe and the assistant probe.

  8. A signal-on electrochemical aptasensor for ultrasensitive detection of endotoxin using three-way DNA junction-aided enzymatic recycling and graphene nanohybrid for amplification.

    PubMed

    Bai, Lijuan; Chai, Yaqin; Pu, Xiaoyun; Yuan, Ruo

    2014-03-01

    Endotoxin, also known as lipopolysaccharide (LPS), is able to induce a strong immune response on its internalization into mammalian cells. To date, aptamer-based biosensors for LPS detection have been rarely reported. This work describes a new signal-on electrochemical aptasensor for the ultrasensitive detection of LPS by combining the three-way DNA hybridization process and nanotechnology-based amplification. With the help of DNA1 (associated with the concentration of target LPS), the capture probe hybridizes with DNA1 and the assistant probe to open its hairpin structure and form a ternary "Y" junction structure. The DNA1 can be released from the structure in the presence of nicking endonuclease to initiate the next hybridization process. Then a great deal of cleaved capture probe produced in the cyclic process can bind with DNA2-nanocomposite, which contains the electroactive toluidine blue (Tb) with the amplification materials graphene (Gra) and gold nanoparticles (AuNPs). Thus, an enhanced electrochemical signal can be easily read out. With the cascade signal amplification, this newly designed protocol provides an ultrasensitive electrochemical detection of LPS down to the femtogram level (8.7 fg mL(-1)) with a linear range of 6 orders of magnitude (from 10 fg mL(-1) to 50 ng mL(-1)). Moreover, the high sensitivity and specificity make this method versatile for the detection of other biomolecules by changing the corresponding sequences of the capture probe and the assistant probe. PMID:24477782

  9. Enhancing the sensitivity of localized surface plasmon resonance (LSPR) biosensors using nanorods and DNA aptamers

    NASA Astrophysics Data System (ADS)

    Chuang, Po-Chun; Liao, Pei-Chen; Chen, Yih-Fan

    2015-03-01

    Localized surface plasmon resonance (LSPR) biosensors have drawn much attention for their promising application in point-of-care diagnostics. While surface plasmon resonance (SPR) biosensing systems have been well developed, LSPR systems have the advantages of simpler and more compact setups. The LSPR peak shifts caused by the binding of molecules to the LSPR substrates, however, are usually smaller than 1 nm if no signal amplification mechanism is used. When using nanoparticles to enhance the sensitivity of LSPR biosensors, because of the short field penetration depth, the nanoparticles should be very close to the LSPR substrate to induce significant shifts in the LSPR peak position. In this study, we used DNA aptamers and gold nanorods to significantly increase the change in the LSPR peak position with the concentration of the target molecules. We have successfully used the proposed mechanism to detect 0.1 nM interferongamma (IFN-γ), a biomarker related to the diagnosis of latent tuberculosis infection. The calibration curves obtained in pure buffers and serum-containing buffers show that accurate detection can be achieved even when the sample is from complex biological fluids such as serum. Because of the enhancement in the sensitivity by the proposed sensing scheme, it is possible to use a low-cost spectrometer to build a LSPR biosensing system.

  10. Ultrasensitive Electrochemical Detection of mRNA Using Branched DNA Amplifiers

    SciTech Connect

    Mao, Xun; Liu, Guodong; Wang, Shengfu; Lin, Yuehe; Zhang, Aiguo; Zhang, Lurong; Ma, Yunqing

    2008-11-01

    We describe here an ultrasensitive electrochemical detection of m RNA protocol without RNA purification and PCR amplification. The new m RNA electrical detection capability is coupled to the amplification feature of branched DNA (bDNA) technology and with the nagnetic beads based electrochemical bioassay.

  11. Carbon nanotube-based lateral flow biosensor for sensitive and rapid detection of DNA sequence.

    PubMed

    Qiu, Wanwei; Xu, Hui; Takalkar, Sunitha; Gurung, Anant S; Liu, Bin; Zheng, Yafeng; Guo, Zebin; Baloda, Meenu; Baryeh, Kwaku; Liu, Guodong

    2015-02-15

    In this article, we describe a carbon nanotube (CNT)-based lateral flow biosensor (LFB) for rapid and sensitive detection of DNA sequence. Amine-modified DNA detection probe was covalently immobilized on the shortened multi-walled carbon nanotubes (MWCNTs) via diimide-activated amidation between the carboxyl groups on the CNT surface and amine groups on the detection DNA probes. Sandwich-type DNA hybridization reactions were performed on the LFB and the captured MWCNTs on test zone and control zone of LFB produced the characteristic black bands, enabling visual detection of DNA sequences. Combining the advantages of lateral flow chromatographic separation with unique physical properties of MWCNT (large surface area), the optimized LFB was capable of detecting of 0.1 nM target DNA without instrumentation. Quantitative detection could be realized by recording the intensity of the test line with the Image J software, and the detection limit of 40 pM was obtained. This detection limit is 12.5 times lower than that of gold nanoparticle (GNP)-based LFB (0.5 nM, Mao et al. Anal. Chem. 2009, 81, 1660-1668). Another important feature is that the preparation of MWCNT-DNA conjugates was robust and the use of MWCNT labels avoided the aggregation of conjugates and tedious preparation time, which were often met in the traditional GNP-based nucleic acid LFB. The applications of MWCNT-based LFB can be extended to visually detect protein biomarkers using MWCNT-antibody conjugates. The MWCNT-based LFB thus open a new door to prepare a new generation of LFB, and shows great promise for in-field and point-of-care diagnosis of genetic diseases and for the detection of infectious agents. PMID:25262062

  12. "Turn off-on" phosphorescent biosensors for detection of DNA based on quantum dots/acridine orange.

    PubMed

    Miao, Yanming; Li, Yuting; Zhang, Zhifeng; Yan, Guiqin; Bi, Yi

    2015-04-15

    A "turn off-on" switch mode was established by using the interaction between acridine orange (AO) and DNA as an input signal and using the room temperature phosphorescence (RTP) reversible change of 3-mercaptopropionic acid (MPA)-capped Mn-doped ZnS quantum dots (QDs) as an output signal in biological fluids. AO was absorbed into the surface of Mn-doped ZnS QDs via electrostatic attraction and, thus, formed a ground-state complex through photoinduced electron transfer (PIET). This complex quenched the phosphorescence of Mn-doped ZnS QDs and then rendered the system into the "turn-off" mode. Along with the addition of DNA and embedded binding with DNA, AO was competitively induced to fall off from the surface of Mn-doped ZnS QDs and embed into the double helix structure of DNA. As a result, the RTP of Mn-doped ZnS QDs was recovered and the system consequently was rendered into "turn-on" mode. In this case, a new biosensor for DNA detection was built and has a detection limit of 0.033mgL(-1) and a detection range from 0.033 to 20mgL(-1). What is more, this kind of biosensor does not require complex pretreatments and is free from the interference from autofluorescence and scattering light. Thus, this biosensor can be used to detect DNA in biological fluids. PMID:25637306

  13. Porous silicon biosensor: current status.

    PubMed

    Dhanekar, Saakshi; Jain, Swati

    2013-03-15

    Biosensing technologies cater to modern day diagnostics and point of care multi-specialty clinics, hospitals and laboratories. Biosensors aggregate the sensitivity of detection methodologies and constitutional selectivity of biomolecules. Endeavors to develop highly sensitive, fast, stable and low cost biosensors have been made possible by extensive and arduous research. Immense research work is going on for detection of molecules using various materials as immobilization substrate and sensing elements. Amongst materials being used as bio-sensing substrates, nano-porous silicon (PS) has amassed attention and gained popularity in recent years. It has captivating and tunable features like ease of fabrication, special optico-physico properties, tailored morphological structure and versatile surface chemistry enhancing its prospects as transducer for fabricating biosensors. The present review describes the fabrication of PS and its biosensing capabilities for detection of various analytes including, but not limited to, glucose, DNA, antibodies, bacteria and viruses. Attention has been consecrated on the various methodologies such as electrical, electrochemical, optical and label free techniques along with the performances of these biosensors. It concludes with some future prospects and challenges of PS based biosensors. PMID:23122704

  14. Electrochemical Quantification of the Antioxidant Capacity of Medicinal Plants Using Biosensors

    PubMed Central

    Rodríguez-Sevilla, Erika; Ramírez-Silva, María-Teresa; Romero-Romo, Mario; Ibarra-Escutia, Pedro; Palomar-Pardavé, Manuel

    2014-01-01

    The working area of a screen-printed electrode, SPE, was modified with the enzyme tyrosinase (Tyr) using different immobilization methods, namely entrapment with water-soluble polyvinyl alcohol (PVA), cross-linking using glutaraldehyde (GA), and cross-linking using GA and human serum albumin (HSA); the resulting electrodes were termed SPE/Tyr/PVA, SPE/Tyr/GA and SPE/Tyr/HSA/GA, respectively. These biosensors were characterized by means of amperometry and EIS techniques. From amperometric evaluations, the apparent Michaelis-Menten constant, Km′, of each biosensor was evaluated while the respective charge transfer resistance, Rct, was assessed from impedance measurements. It was found that the SPE/Tyr/GA had the smallest Km′ (57 ± 7) μM and Rct values. This electrode also displayed both the lowest detection and quantification limits for catechol quantification. Using the SPE/Tyr/GA, the Trolox Equivalent Antioxidant Capacity (TEAC) was determined from infusions prepared with “mirto” (Salvia microphylla), “hHierba dulce” (Lippia dulcis) and “salve real” (Lippia alba), medicinal plants commonly used in Mexico. PMID:25111237

  15. Electrochemical sensors and biosensors for determination of catecholamine neurotransmitters: A review.

    PubMed

    Ribeiro, José A; Fernandes, Paula M V; Pereira, Carlos M; Silva, F

    2016-11-01

    This work describes the state of the art of electrochemical devices for the detection of an important class of neurotransmitters: the catecholamines. This class of biogenic amines includes dopamine, noradrenaline (also called norepinephrine) and adrenaline (also called epinephrine). Researchers have focused on the role of catecholamine molecules within the human body because they are involved in many important biological functions and are commonly associated with several diseases, such as Alzheimer's and Parkinson. Furthermore, the release of catecholamines as a consequence of induced stimulus is an important indicator of reward-related behaviors, such as food, drink, sex and drug addiction. Thus, the development of simple, fast and sensitive electroanalytical methodologies for the determination of catecholamines is currently needed in clinical and biomedical fields, as they have the potential to serve as clinically relevant biomarkers for specific disease states or to monitor treatment efficacy. Currently, three main strategies have used by researchers to detect catecholamine molecules, namely: the use electrochemical materials in combination with, for example, HPLC or FIA, the incorporation of new materials/layers on the sensor surfaces (Tables 1-7) and in vivo detection, manly by using FSCV at CFMEs (Section 10). The developed methodologies were able not only to accurately detect catecholamines at relevant concentration levels, but to do so in the presence of co-existing interferences in samples detected (ascorbate, for example). This review examines the progress made in electrochemical sensors for the selective detection of catecholamines in the last 15 years, with special focus on highly innovative features introduced by nanotechnology. As the literature in rather extensive, we try to simplify this work by summarizing and grouping electrochemical sensors according to the manner their substrates were chemically modified. We also discuss the current and future

  16. Simultaneous measurement of cholinergic tone and neuronal network dynamics in vivo in the rat brain using a novel choline oxidase based electrochemical biosensor.

    PubMed

    Santos, Ricardo M; Laranjinha, João; Barbosa, Rui M; Sirota, Anton

    2015-07-15

    Acetylcholine (ACh) modulates neuronal network activities implicated in cognition, including theta and gamma oscillations but the mechanisms remain poorly understood. Joint measurements of cholinergic activity and neuronal network dynamics with high spatio-temporal resolution are critical to understand ACh neuromodulation. However, current electrochemical biosensors are not optimized to measure nanomolar cholinergic signals across small regions like hippocampal sub-layers. Here, we report a novel oxidase-based electrochemical biosensor that matches these constraints. The approach is based on measurement of H2O2 generated by choline oxidase (ChOx) in the presence of choline (Ch). The microelectrode design consists of a twisted pair of 50µm diameter Pt/Ir wires (sensor and sentinel), which is scalable, provides high spatial resolution and optimizes common mode rejection. Microelectrode coating with ChOx in chitosan cross-linked with benzoquinone is simple, mechanically robust and provides high sensitivity (324±46nAµM(-1)cm(-2)), a limit of detection of 16nM and a t50 response time of 1.4s. Local field potential (LFP)-related currents dominate high-frequency component of electrochemical recordings in vivo. We significantly improved signal-to-noise-ratio compared to traditional sentinel subtraction by a novel frequency domain common mode rejection procedure that accounts for differential phase and amplitude of LFP-related currents on the two channels. We demonstrate measurements of spontaneous nanomolar Ch fluctuations, on top of which micromolar Ch increases occurred during periods of theta activity in anesthetized rats. Measurements were not affected by physiological O2 changes, in agreement with the low biosensor Km for O2 (2.6µM). Design and performance of the novel biosensor opens the way for multisite recordings of spontaneous cholinergic dynamics in behaving animals. PMID:25706061

  17. Integrating SPR-ellipsometry and electrochemical measurements for performance evaluation of label-free thiophene-based biosensor

    NASA Astrophysics Data System (ADS)

    Tsai, Pei-I.; Lee, Shu-Sheng; Chou, Shin-Ting; Chang, Yu-Ting; Lee, Adam Shih-Yuan; Lee, C. K.

    2014-03-01

    The surface plasmon resonance reflectance changes measured with a circularly polarized ellipsometry and an electrochemical impedance spectroscopy were identified to be able to characterize the critical roles of biomolecules for vastly different biological functions and processes. Throughout the course of this study, interferon-gamma (IFN-γ) was chosen as the biomarker to test and to verify the performance of this newly developed system for Tuberculosis detection. The interactions of IFN-γ with immobilized anti-IFN-γ antibody at various concentrations were interrogated both optically and electrochemically. A semi-conductive linker bis-thiophene was thiolated to ensure the cross-linked monoclonal human IFN-γ antibody got self-assembled onto the gold thin film and form a label-free biosensor. The functional features of the bis-thiophene coated-gold film were characterized by cyclic voltammetry and impedance spectroscopy methods. The association of IFN-γ to the bis-thiophene bridging units via antibody-antigen interactions provided the basis for ultrasensitive detection of IFN-γ by tracking the conformation changes in surface-bound protein molecules. The phase shift can be attributed to the average thickness and the real-time index of refraction of the protein layer in different protein layer. Experimental results obtained by impedance spectroscopy and by phase-interrogation SPR showed linear dynamic range. Our experimental results verified that an increase in the concentration of the IFN-γ usually accompanied by phase increase in SPR and an impedance decrease in EIS. These results indicated that our newly developed integrated biosensing system can potentially provide new insight into various conjugate phenomena and interfacial processes for observing molecular conformation changes.

  18. A reusable optical biosensor for the ultrasensitive and selective detection of unamplified human genomic DNA with gold nanostars.

    PubMed

    Mariani, Stefano; Scarano, Simona; Spadavecchia, Jolanda; Minunni, Maria

    2015-12-15

    A Surface Plasmon Resonance imaging (SPRi) based DNA sensors for the selective and ultrasensitive human genomic DNA detection, directly extracted from lymphocytes (bypassing PCR amplification), is reported. To achieve DNA detection, a rationally chosen star-shaped nanoparticle (NP), namely gold nanostar (AuNS), has been applied, for the first time, in a sandwich-like assay based on the selective capturing of specific DNA targets and the subsequent signal amplification by a secondary DNA probe linked to AuNS. The plasmonic profile, size and electric field enhancements at the star tips contributed to the maximization of plasmon coupling between LSPs and SPs as aimed for analytical signal magnification. The system was first tested using short synthetic DNA target sequences and applied to DNA biosensing, lowering 610-fold the detection limit from 6.1 nM (without NSs labeling) to 10 pM (with NSs labeling). Then the biosensor was applied to genomic DNA samples, extracted from human lymphocytes and undergoing only to a simple ultrasonic fragmentation, lowering (~435 fold) the detection limit from 3.0 fM (without NSs labeling) to 6.9 aM (with NSs labeling). Thanks to the assay optimization, we proved that tuning the NSs surface coverage with DNA linked to nanoparticles is crucial not only for the increase of signals but also for the regenerability/reusability of the biosensor for tens of measurement cycles. PMID:26264264

  19. Shear acoustic wave biosensor for detecting DNA intrinsic viscosity and conformation: a study with QCM-D.

    PubMed

    Tsortos, Achilleas; Papadakis, George; Gizeli, Electra

    2008-12-01

    Direct biosensors are devices operating by monitoring the amount of surface-bound analyte. In this work a new approach is presented where a label-free acoustic biosensor, based on a QCM-D device, and solution viscosity theory, are used to study DNA intrinsic viscosity. The latter is quantitatively related to the DNA conformation and specifically the molecule's shape and size, in a manner that is independent of the amount of bound DNA mass. It is shown that acoustic measurements can clearly distinguish between ds-DNA of same shape (straight rod) but various sizes (from 20 to 198bp (base pairs)) and same mass and size (90bp) but various shapes ("straight", "bent", "triangle"). These results are discussed in the broader context of "coil" and sphere-like molecules detected on surfaces. A mathematical formula is presented relating the length of straight, surface-protruding DNA to the acoustic ratio DeltaD/Deltaf. The development of real-time rapid techniques for the characterization of DNA intrinsic curvature as well as DNA conformational changes upon interaction with proteins is of significance to analytical biotechnology due to the large number of DNA sequences and potential DNA bending proteins involved in genome analysis and drug screening. PMID:18723337

  20. Glutaraldehyde activated eggshell membrane for immobilization of tyrosinase from Amorphophallus companulatus: application in construction of electrochemical biosensor for dopamine.

    PubMed

    Tembe, Sanket; Kubal, B S; Karve, Meena; D'Souza, S F

    2008-04-01

    Tyrosinase from a plant source Amorphophallus companulatus was immobilized on eggshell membrane using glutaraldehyde. Among the three different approaches used for immobilization, activation of eggshell membrane by glutaraldehyde followed by enzyme adsorption on activated support could stabilize the enzyme tyrosinase and was found to be effective. K(m) and V(max) values for dopamine hydrochloride calculated from Lineweaver-Burk plot were 0.67 mM and 0.08 mM min(-1), respectively. Studies on effect of pH showed retention of more than 90% activity over a pH range 5.0-6.5. Membrane bound enzyme exhibited consistent activity in the temperature range 20-45 degrees C. Shelf life of immobilized tyrosinase system was found to be more than 6 months when stored in phosphate buffer at 4 degrees C. An electrochemical biosensor for dopamine was developed by mounting the tyrosinase immobilized eggshell membrane on the surface of glassy carbon electrode. Dopamine concentrations were determined by the direct reduction of biocatalytically liberated quinone species at -0.19 V versus Ag/AgCl (3M KCl). Linearity was observed within the range of 50-250 microM with a detection limit of 25 microM. PMID:18358868

  1. DNA aptamer-based fiber optic biosensor for selective and label-free detection of dopamine

    NASA Astrophysics Data System (ADS)

    Zibaii, M. I.; Latifi, H.; Asadollahi, A.; Bayat, A. H.; Haghparast, A.

    2015-09-01

    Dopamine (DA) analysis is complicated by the interference from other electrochemically active endogenous compounds present in the brain, including DA precursors and metabolites and other neurotransmitters (NT). Here we report a simple, sensitive and selective optical fiber biosensor for the detection of DA in the presence of other NT. It is composed of a 57-mer dopamine-binding aptamer (DBA) as recognition element and nonadiabatic tapered optical fiber (NATOF) as probe. Upon the addition of DA, the conformation of DBA would change from a random coil structure to a rigid tertiary structure like a pocket. The conformational change of DBA lead to the refractive index (RI) change around the tapered fiber surface. Specific recognition of DA by the aptamer allowed a selective optical detection of DA within the physiologically relevant 500 nM to 10 μM range. Some common interferents such as epinephrine (EP) and ascorbic acid (AA) showed no or just a little interference in the determination of DA.

  2. Architecture of a modular, multichannel readout system for dense electrochemical biosensor microarrays

    NASA Astrophysics Data System (ADS)

    Ramfos, Ioannis; Blionas, Spyridon; Birbas, Alexios

    2015-01-01

    The architecture of a modular, multichannel readout system for dense electrochemical microarrays, targeting Lab-on-a-Chip applications, is presented. This approach promotes efficient component reusability through a hybrid multiplexing methodology, maintaining high levels of sampling performance and accuracy. Two readout modes are offered, which can be dynamically interchanged following signal profiling, to cater for both rapid signal transitions and weak current responses. Additionally, functional extensions to the described architecture are discussed, which provide the system with multi-biasing capabilities. A prototype integrated circuit of the proposed architecture’s analog core and a supporting board were implemented to verify the working principles. The system was evaluated using standard loads, as well as electrochemical sensor arrays. Through a range of operating conditions and loads, the prototype exhibited a highly linear response and accurately delivered the readout of input signals with fast transitions and wide dynamic ranges.

  3. Interface Design for CMOS-Integrated Electrochemical Impedance Spectroscopy (EIS) Biosensors

    PubMed Central

    Manickam, Arun; Johnson, Christopher Andrew; Kavusi, Sam; Hassibi, Arjang

    2012-01-01

    Electrochemical Impedance Spectroscopy (EIS) is a powerful electrochemical technique to detect biomolecules. EIS has the potential of carrying out label-free and real-time detection, and in addition, can be easily implemented using electronic integrated circuits (ICs) that are built through standard semiconductor fabrication processes. This paper focuses on the various design and optimization aspects of EIS ICs, particularly the bio-to-semiconductor interface design. We discuss, in detail, considerations such as the choice of the electrode surface in view of IC manufacturing, surface linkers, and development of optimal bio-molecular detection protocols. We also report experimental results, using both macro- and micro-electrodes to demonstrate the design trade-offs and ultimately validate our optimization procedures. PMID:23202170

  4. Electrochemical Glucose Sensors—Developments Using Electrostatic Assembly and Carbon Nanotubes for Biosensor Construction

    PubMed Central

    Harper, Alice; Anderson, Mark R.

    2010-01-01

    In 1962, Clark and Lyons proposed incorporating the enzyme glucose oxidase in the construction of an electrochemical sensor for glucose in blood plasma. In their application, Clark and Lyons describe an electrode in which a membrane permeable to glucose traps a small volume of solution containing the enzyme adjacent to a pH electrode, and the presence of glucose is detected by the change in the electrode potential that occurs when glucose reacts with the enzyme in this volume of solution. Although described nearly 50 years ago, this seminal development provides the general structure for constructing electrochemical glucose sensors that is still used today. Despite the maturity of the field, new developments that explore solutions to the fundamental limitations of electrochemical glucose sensors continue to emerge. Here we discuss two developments of the last 15 years; confining the enzyme and a redox mediator to a very thin molecular films at electrode surfaces by electrostatic assembly, and the use of electrodes modified by carbon nanotubes (CNTs) to leverage the electrocatalytic effect of the CNTs to reduce the oxidation overpotential of the electrode reaction or for the direct electron transport to the enzyme. PMID:22163652

  5. Electrochemical Biosensor Based on Nanoporous Au/CoO Core-Shell Material with Synergistic Catalysis.

    PubMed

    Zhang, Chao; Huang, Bin; Qian, Lihua; Yuan, Songliu; Wang, Shuai; Chen, Rong

    2016-01-01

    An ultrathin CoO layer is deposited on the skeleton surfaces of a nanoporous gold (NPG) film by using atomic layer deposition, creating a flexible electrode. Detailed characterization demonstrates the superior performance of the flexible NPG/CoO hybrids for electrochemical catalysis. The NPG/CoO hybrid not only achieves high catalytic activity for glucose oxidation and H2O2 reduction, but also exhibits a linear dependence of the electrical signal on the concentration of glucose and H2O2 molecules in the electrolyte. Meanwhile, the sensitivity for H2O2 reduction can be as high as 62.5 μA mm(-1)  cm(-2) with linear dependence on the concentration in the range of 0.1-92.9 mm. The high sensitivity is proposed to result from the synergistic effect of Au and CoO at the interfaces, and the high conductivity of the gold skeleton with a large surface area. The superior electrochemical performance of this hybrid electrode is promising for future potential applications in various transitional-metal-oxide-based electrochemical electrodes. PMID:26521684

  6. Gold nanoparticles-induced enhancement of the analytical response of an electrochemical biosensor based on an organic-inorganic hybrid composite material.

    PubMed

    Barbadillo, M; Casero, E; Petit-Domínguez, M D; Vázquez, L; Pariente, F; Lorenzo, E

    2009-12-15

    The design and characterization of a new organic-inorganic hybrid composite material for glucose electrochemical sensing are described. This material is based on the entrapment of both gold nanoparticles (AuNPs) and glucose oxidase, which was chosen as a model, into a sol-gel matrix. The addition of spectroscopic grade graphite to this system, which confers conductivity, leads to the development of a material particularly attractive for electrochemical biosensor fabrication. The characterization of the hybrid composite material was performed using atomic force microscopy and scanning electron microscopy techniques. This composite material was applied to the determination of glucose in presence of hydroxymethylferrocene as a redox mediator. The system exhibits a clear electrocatalytic activity towards glucose, allowing its determination at 250 mV vs Ag/AgCl. The performance of the resulting enzyme biosensor was evaluated in terms of sensitivity, detection limit, linear response range, stability and accuracy. Finally, the enhancement of the analytical response of the resulting biosensor induced by the presence of gold nanoparticles was evaluated by comparison with a similar organic-inorganic hybrid composite material without AuNPs. PMID:19836554

  7. Electrochemical detection of harmful algae and other microbial contaminants in coastal waters using hand-held biosensors.

    PubMed

    LaGier, Michael J; Fell, Jack W; Goodwin, Kelly D

    2007-06-01

    Standard methods to identify microbial contaminants in the environment are slow, laborious, and can require specialized expertise. This study investigated electrochemical detection of microbial contaminants using commercially available, hand-held instruments. Electrochemical assays were developed for a red tide dinoflagellate (Karenia brevis), fecal-indicating bacteria (Enterococcus spp.), markers indicative of human sources of fecal pollution (human cluster Bacteroides and the esp gene of Enterococcus faecium), bacterial pathogens (Escherichia coli 0157:H7, Salmonella spp., Campylobacter jejuni, Staphylococcus aureus), and a viral pathogen (adenovirus). For K. brevis, two assay formats (Rapid PCR-Detect and Hybrid PCR-Detect) were tested and both provided detection limits of 10 genome equivalents for DNA isolated from K. brevis culture and amplified by PCR. Sensitivity with coastal water samples was sufficient to detect K. brevis that was "present" (electrochemical signal was significantly different than for samples containing cells at "medium" concentrations (100,000 to<10(6)cells/l). Detection of K. brevis RNA was also shown. Multi-target capability was demonstrated with an 8-plex assay for bacterial and viral targets using isolated DNA, natural beach water spiked with human feces, and water and sediments collected from New Orleans, Louisiana following Hurricane Katrina. Furthermore, direct detection of dinoflagellate and bacterial DNA was achieved using lysed cells rather than extracted nucleic acids, allowing streamlining of the process. The methods presented can be used to rapidly (3-5h) screen environmental water samples for the presence of microbial contaminants and have the potential to be integrated into semi-automated detection platforms. PMID:17328925

  8. Development of mercury (II) ion biosensors based on mercury-specific oligonucleotide probes.

    PubMed

    Li, Lanying; Wen, Yanli; Xu, Li; Xu, Qin; Song, Shiping; Zuo, Xiaolei; Yan, Juan; Zhang, Weijia; Liu, Gang

    2016-01-15

    Mercury (II) ion (Hg(2+)) contamination can be accumulated along the food chain and cause serious threat to the public health. Plenty of research effort thus has been devoted to the development of fast, sensitive and selective biosensors for monitoring Hg(2+). Thymine was demonstrated to specifically combine with Hg(2+) and form a thymine-Hg(2+)-thymine (T-Hg(2+)-T) structure, with binding constant even higher than T-A Watson-Crick pair in DNA duplex. Recently, various novel Hg(2+) biosensors have been developed based on T-rich Mercury-Specific Oligonucleotide (MSO) probes, and exhibited advanced selectivity and excellent sensitivity for Hg(2+) detection. In this review, we explained recent development of MSO-based Hg(2+) biosensors mainly in 3 groups: fluorescent biosensors, colorimetric biosensors and electrochemical biosensors. PMID:26356764

  9. In vivo Electrochemical Biosensor for Brain Glutamate Detection: A Mini Review.

    PubMed

    Hamdan, Siti Kartika; Mohd Zain, Ainiharyati

    2014-12-01

    Glutamate is one of the most prominent neurotransmitters in mammalian brains, which plays an important role in neuronal excitation. High levels of neurotransmitter cause numerous alterations, such as calcium overload and the dysfunction of mitochondrial and oxidative stress. These alterations may lead to excitotoxicity and may trigger multiple neuronal diseases, such as Alzheimer's disease, stroke, and epilepsy. Excitotoxicity is a pathological process that damages nerve cells and kills cells via excessive stimulation by neurotransmitters. Monitoring the concentration level of brain glutamate via an implantable microbiosensor is a promising alternative approach to closely investigate in the function of glutamate as a neurotransmitter. This review outlines glutamate microbiosensor designs to enhance the sensitivity of glutamate detection with less biofouling occurrence and minimal detection of interference species. There are many challenges in the development of a reproducible and stable implantable microbiosensor because many factors and limitations may affect the detection performance. However, the incorporation of multiple scales is needed to address the basic issues and combinations across the various disciplines needed to achieve the success of the system to overcome the challenges in the development of an implantable glutamate biosensor. PMID:25941459

  10. In vivo Electrochemical Biosensor for Brain Glutamate Detection: A Mini Review

    PubMed Central

    HAMDAN, Siti Kartika; MOHD ZAIN, ainiharyati

    2014-01-01

    Glutamate is one of the most prominent neurotransmitters in mammalian brains, which plays an important role in neuronal excitation. High levels of neurotransmitter cause numerous alterations, such as calcium overload and the dysfunction of mitochondrial and oxidative stress. These alterations may lead to excitotoxicity and may trigger multiple neuronal diseases, such as Alzheimer’s disease, stroke, and epilepsy. Excitotoxicity is a pathological process that damages nerve cells and kills cells via excessive stimulation by neurotransmitters. Monitoring the concentration level of brain glutamate via an implantable microbiosensor is a promising alternative approach to closely investigate in the function of glutamate as a neurotransmitter. This review outlines glutamate microbiosensor designs to enhance the sensitivity of glutamate detection with less biofouling occurrence and minimal detection of interference species. There are many challenges in the development of a reproducible and stable implantable microbiosensor because many factors and limitations may affect the detection performance. However, the incorporation of multiple scales is needed to address the basic issues and combinations across the various disciplines needed to achieve the success of the system to overcome the challenges in the development of an implantable glutamate biosensor. PMID:25941459

  11. Improvement of Electrochemical Response of Cocaine Sensors Based on DNA Aptamer by Heat Treatment.

    PubMed

    Arimoto, Satoshi; Shimono, Ken; Yasukawa, Tomoyuki; Mizutani, Fumio; Yoshioka, Toshihiko

    2016-01-01

    We report on a biosensor for cocaine based on the conformation change of DNA aptamer by capturing the cocaine molecules. The oxidation current of ferrocene conjugated on the terminal end of aptamer immobilized on an Au electrode increased with increasing cocaine concentration. The sensor response has been improved by a simple heat treatment after immobilization, since the aggregates of DNA aptamer generated during the immobilization step could be dissociated and rearranged on the electrode. PMID:27063722

  12. Porphyrin-encapsulated metal-organic frameworks as mimetic catalysts for electrochemical DNA sensing via allosteric switch of hairpin DNA.

    PubMed

    Ling, Pinghua; Lei, Jianping; Zhang, Lei; Ju, Huangxian

    2015-04-01

    A sensitive electrochemical sensor is designed for DNA detection based on mimetic catalysis of metal-organic framework (MOF) and allosteric switch of hairpin DNA. The functional MOFs are synthesized as signal probes by a one-pot encapsulation of iron(III) meso-5,10,15,20-tetrakis(4-carboxyphenyl) porphyrin chloride (FeTCPP) into a prototypal MOF, HKUST-1(Cu), and sequentially conjugated with streptavidin (SA) as a recognition element. The resulting FeTCPP@MOF composites can mimetically catalyze the oxidation of o-phenylenediamine (o-PD) to 2,2'-diaminoazobenzene, which is a good electrochemical indicator for signal readout. The presence of target DNA introduces the allosteric switch of hairpin DNA to form SA aptamer, and thus, FeTCPP@MOF-SA probe is brought on the electrode surface via the specific recognition between SA and the corresponding aptamer, resulting in the enhancement of electrochemical signal. The "signal-on" electrochemical sensor can detect target DNA down to 0.48 fM with the linear range of 10 fM to 10 nM. Moreover, the MOF-based electrochemical sensor exhibits acceptable selectivity against even a single mismatched DNA and good feasibility in complex serum matrixes. This strategy opens up a new direction of porphyrin-functionalized MOF for signal transduction in electrochemical biosensing. PMID:25741988

  13. DNA biosensors based on layer-by-layer self-assembled multilayer films of carbon nanotubes and gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Xiao, Yiyun; Dai, Zhao; Zhang, Jimei; Pang, Jiechun; Xu, Shichao; Zheng, Guo

    2009-07-01

    A novel DNA biosensor based on layer-by-layer self-assembled multi-walled carbon nanotubes (MWNTs) and gold nano-particles (GNPs) was presented in this paper, in which the probe HS-ssDNA oligonucleotides, MWNTs and GNPs were all covalently immobilized by chemical Au-Sulphide bonding. Firstly, the super short MWNTs were prepared and modified with thio groups which could be self-assembled onto the surface of Au elcetrode by Au-sulphide bonding, then the GNPs were chemically adhered to the surfaces of MWNTs by forming Au-sulphide bonding again, at last the selfassamble of probe DNA oligonucleotides were also covalently immobilized via Au-sulphide bonding between thio groups at the ends of the DNA oligonucleotides and GNPs. Hybridization between the probe HS-ssDNA oligonucleotides and target DNA oligonucleotides was confirmed by the changes in the voltammetric peak of an anionic intercalator, anthraquinone-2,6-disulfonic acid (AQDS) as a hybridization indicator. The cyclic voltammetric and differential pulse voltammetry responses demonstrated that the DNA biosensors based on Layer-by-layer self-assembled multilayer films of MWNTs and NGPs offer a higher hybridization efficiency and selectivity compared to those based on only random MWNTs or GNPs.

  14. Electrochemical Patterning and Detection of DNA Arrays on a Two-Electrode Platform

    PubMed Central

    Furst, Ariel; Landefeld, Sally; Hill, Michael G.; Barton, Jacqueline K.

    2014-01-01

    We report a novel method of DNA array formation that is electrochemically formed and addressed with a two-electrode platform. Electrochemical activation of a copper catalyst, patterned with one electrode, enables precise placement of multiple sequences of DNA onto a second electrode surface. The two-electrode patterning and detection platform allows for both spatial resolution of the patterned DNA array and optimization of detection through DNA-mediated charge transport with electrocatalysis. This two-electrode platform has been used to form arrays that enable differentiation between well-matched and mismatched sequences, the detection of TATA-binding protein, and sequence-selective DNA hybridization. PMID:24328227

  15. Electrochemical patterning and detection of DNA arrays on a two-electrode platform.

    PubMed

    Furst, Ariel; Landefeld, Sally; Hill, Michael G; Barton, Jacqueline K

    2013-12-26

    We report a novel method of DNA array formation that is electrochemically formed and addressed with a two-electrode platform. Electrochemical activation of a copper catalyst, patterned with one electrode, enables precise placement of multiple sequences of DNA onto a second electrode surface. The two-electrode patterning and detection platform allows for both spatial resolution of the patterned DNA array and optimization of detection through DNA-mediated charge transport with electrocatalysis. This two-electrode platform has been used to form arrays that enable differentiation between well-matched and mismatched sequences, the detection of TATA-binding protein, and sequence-selective DNA hybridization. PMID:24328227

  16. DNA Nanostructure-based Interfacial engineering for PCR-free ultrasensitive electrochemical analysis of microRNA

    NASA Astrophysics Data System (ADS)

    Wen, Yanli; Pei, Hao; Shen, Ye; Xi, Junjie; Lin, Meihua; Lu, Na; Shen, Xizhong; Li, Jiong; Fan, Chunhai

    2012-11-01

    MicroRNAs (miRNAs) have been identified as promising cancer biomarkers due to their stable presence in serum. As an alternative to PCR-based homogenous assays, surface-based electrochemical biosensors offer great opportunities for low-cost, point-of-care tests (POCTs) of disease-associated miRNAs. Nevertheless, the sensitivity of miRNA sensors is often limited by mass transport and crowding effects at the water-electrode interface. To address such challenges, we herein report a DNA nanostructure-based interfacial engineering approach to enhance binding recognition at the gold electrode surface and drastically improve the detection sensitivity. By employing this novel strategy, we can directly detect as few as attomolar (<1, 000 copies) miRNAs with high single-base discrimination ability. Given that this ultrasensitive electrochemical miRNA sensor (EMRS) is highly reproducible and essentially free of prior target labeling and PCR amplification, we also demonstrate its application by analyzing miRNA expression levels in clinical samples from esophageal squamous cell carcinoma (ESCC) patients.

  17. DNA Nanostructure-based Interfacial engineering for PCR-free ultrasensitive electrochemical analysis of microRNA

    PubMed Central

    Wen, Yanli; Pei, Hao; Shen, Ye; Xi, Junjie; Lin, Meihua; Lu, Na; Shen, Xizhong; Li, Jiong; Fan, Chunhai

    2012-01-01

    MicroRNAs (miRNAs) have been identified as promising cancer biomarkers due to their stable presence in serum. As an alternative to PCR-based homogenous assays, surface-based electrochemical biosensors offer great opportunities for low-cost, point-of-care tests (POCTs) of disease-associated miRNAs. Nevertheless, the sensitivity of miRNA sensors is often limited by mass transport and crowding effects at the water-electrode interface. To address such challenges, we herein report a DNA nanostructure-based interfacial engineering approach to enhance binding recognition at the gold electrode surface and drastically improve the detection sensitivity. By employing this novel strategy, we can directly detect as few as attomolar (<1, 000 copies) miRNAs with high single-base discrimination ability. Given that this ultrasensitive electrochemical miRNA sensor (EMRS) is highly reproducible and essentially free of prior target labeling and PCR amplification, we also demonstrate its application by analyzing miRNA expression levels in clinical samples from esophageal squamous cell carcinoma (ESCC) patients. PMID:23162691

  18. Molecularly imprinted electrochemical biosensor based on Fe@Au nanoparticles involved in 2-aminoethanethiol functionalized multi-walled carbon nanotubes for sensitive determination of cefexime in human plasma.

    PubMed

    Yola, Mehmet Lütfi; Eren, Tanju; Atar, Necip

    2014-10-15

    The molecular imprinting technique depends on the molecular recognition. It is a polymerization method around the target molecule. Hence, this technique creates specific cavities in the cross-linked polymeric matrices. In present study, a sensitive imprinted electrochemical biosensor based on Fe@Au nanoparticles (Fe@AuNPs) involved in 2-aminoethanethiol (2-AET) functionalized multi-walled carbon nanotubes (f-MWCNs) modified glassy carbon (GC) electrode was developed for determination of cefexime (CEF). The results of X-ray photoelectron spectroscopy (XPS) and reflection-absorption infrared spectroscopy (RAIRS) confirmed the formation of the developed surfaces. CEF imprinted film was constructed by cyclic voltammetry (CV) for 9 cycles in the presence of 80 mM pyrrole in phosphate buffer solution (pH 6.0) containing 20mM CEF. The developed electrochemical biosensor was validated according to the International Conference on Harmonisation (ICH) guideline and found to be linear, sensitive, selective, precise and accurate. The linearity range and the detection limit were obtained as 1.0 × 10(-10)-1.0 × 10(-8)M and 2.2 × 10(-11)M, respectively. The developed CEF imprinted sensor was successfully applied to real samples such as human plasma. In addition, the stability and reproducibility of the prepared molecular imprinted electrode were investigated. The excellent long-term stability and reproducibility of the prepared CEF imprinted electrodes make them attractive in electrochemical sensors. PMID:24832202

  19. A novel electrically magnetic-controllable electrochemical biosensor for the ultra sensitive and specific detection of attomolar level oral cancer-related microRNA.

    PubMed

    Wang, ZongWen; Zhang, Jing; Guo, Ying; Wu, XiaoYan; Yang, WeiJuan; Xu, LiangJun; Chen, JingHua; Fu, FengFu

    2013-07-15

    Non-invasive early diagnosis of oral cancer is the most effective means to reduce mortality rate from this disease. In this paper, we described a novel magnetic-controllable electrochemical RNA biosensor for the ultra sensitive and specific detection of oral cancer-related microRNA (miRNA) based on a home-made electrically magnetic-controllable gold electrode. The electrically magnetic-controllable gold electrode combined the merits of heated electrode and magnetic electrode, has notable advantage such as that the strength and direction of the magnetic field and the temperature of the electrode's surface can be easily regulated. The advantage of electrically magnetic-controllable gold electrode, as well as the utilization of "junction-probe" strategy and magnetic beads (MBs)-based enzymatic catalysis amplification, make the biosensor has ultra-high sensitivity and discrimination ability even for the detection of similar miRNAs. It can be used to detect as low as 0.22 aM (2.2 × 10(-19)M) of oral cancer-related miRNA with a recovery of 93-108% and a RSD<6 (n=5). The high sensitivity and selectivity, as well as the easiness of fabrication, operational convenience, short analysis time, good stability and re-usability, make the biosensor a promising alternative for the early point-of-care diagnosis of oral cancer. The success of the biosensor also leads to a great potential in the development of biosensor for the early diagnosis of other diseases. PMID:23455049

  20. Electrical detection of dengue virus (DENV) DNA oligomer using silicon nanowire biosensor with novel molecular gate control.

    PubMed

    Nuzaihan M N, M; Hashim, U; Md Arshad, M K; Kasjoo, S R; Rahman, S F A; Ruslinda, A R; Fathil, M F M; Adzhri, R; Shahimin, M M

    2016-09-15

    In this paper, a silicon nanowire biosensor with novel molecular gate control has been demonstrated for Deoxyribonucleic acid (DNA) detection related to dengue virus (DENV). The silicon nanowire was fabricated using the top-down nanolithography approach, through nanostructuring of silicon-on-insulator (SOI) layers achieved by combination of the electron-beam lithography (EBL), plasma dry etching and size reduction processes. The surface of the fabricated silicon nanowire was functionalized by means of a three-step procedure involving surface modification, DNA immobilization and hybridization. This procedure acts as a molecular gate control to establish the electrical detection for 27-mers base targets DENV DNA oligomer. The electrical detection is based on the changes in current, resistance and conductance of the sensor due to accumulation of negative charges added by the immobilized probe DNA and hybridized target DNA. The sensitivity of the silicon nanowire biosensors attained was 45.0µAM(-1), which shows a wide-range detection capability of the sensor with respect to DNA. The limit of detection (LOD) achieved was approximately 2.0fM. The demonstrated results show that the silicon nanowire has excellent properties for detection of DENV with outstanding repeatability and reproducibility performances. PMID:27107147

  1. A regenerated electrochemical biosensor for label-free detection of glucose and urea based on conformational switch of i-motif oligonucleotide probe.

    PubMed

    Gao, Zhong Feng; Chen, Dong Mei; Lei, Jing Lei; Luo, Hong Qun; Li, Nian Bing

    2015-10-15

    Improving the reproducibility of electrochemical signal remains a great challenge over the past decades. In this work, i-motif oligonucleotide probe-based electrochemical DNA (E-DNA) sensor is introduced for the first time as a regenerated sensing platform, which enhances the reproducibility of electrochemical signal, for label-free detection of glucose and urea. The addition of glucose or urea is able to activate glucose oxidase-catalyzed or urease-catalyzed reaction, inducing or destroying the formation of i-motif oligonucleotide probe. The conformational switch of oligonucleotide probe can be recorded by electrochemical impedance spectroscopy. Thus, the difference of electron transfer resistance is utilized for the quantitative determination of glucose and urea. We further demonstrate that the E-DNA sensor exhibits high selectivity, excellent stability, and remarkable regenerated ability. The human serum analysis indicates that this simple and regenerated strategy holds promising potential in future biosensing applications. PMID:26515000

  2. Electrochemical investigation of interaction between mitomycin C and DNA in a novel drug-delivery system.

    PubMed

    Karadeniz, Hakan; Alparslan, Levent; Erdem, Arzum; Karasulu, Ercument

    2007-10-18

    A novel drug-delivery system was developed by loading the anticancer drug, mitomycin C (MC) into an oil/water system with the aim of investigation by electrochemical sensing the interaction between the drug and DNA in microemulsion phase. The physical and physicochemical properties (droplet size, pH, viscosity, conductivity and refractive index) of this microemulsion were examined. The electrochemical detection of the interaction between MC and double-stranded DNA (dsDNA) in microemulsion phase was performed by using differential pulse voltammetry (DPV) in combination with a disposable sensor, pencil graphite electrode (PGE). The magnitude of guanine oxidation signal was monitored before and after interaction between MC and dsDNA. The effect of different experimental parameters, such as MC concentration, MC interaction time with dsDNA, and dsDNA concentration were also studied to find the optimum analytical performance based on electrochemical detection of this interaction in microemulsion phase. PMID:17590560

  3. New CNT/poly(brilliant green) and CNT/poly(3,4-ethylenedioxythiophene) based electrochemical enzyme biosensors.

    PubMed

    Barsan, Madalina M; Pifferi, Valentina; Falciola, Luigi; Brett, Christopher M A

    2016-07-13

    A combination of the electroactive polymer poly(brilliant green) (PBG) or conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) with carbon nanotubes to obtain CNT/PBG and CNT/PEDOT modified carbon film electrodes (CFE) has been investigated as a new biosensor platform, incorporating the enzymes glucose oxidase (GOx) as test enzyme, alcohol oxidase (AlcOx) or alcohol dehydrogenase (AlcDH). The sensing parameters were optimized for all biosensors based on CNT/PBG/CFE, CNT/PEDOT/CFE platforms. Under optimized conditions, both GOx biosensors exhibited very similar sensitivities, while in the case of AlcOx and AlcDH biosensors, AlcOx/CNT/PBG/CFE was found to give a higher sensitivity and lower detection limit. The influence of dissolved O2 on oxidase-biosensor performance was investigated and was shown to be different for each enzyme. Comparisons were made with similar reported biosensors, showing the advantages of the new biosensors, and excellent selectivity against potential interferents was successfully demonstrated. Finally, alcohol biosensors were successfully used for the determination of ethanol in alcoholic beverages. PMID:27237835

  4. Development of an electrochemical biosensor for the detection of aflatoxin M1 in milk.

    PubMed

    Paniel, Nathalie; Radoi, Antonio; Marty, Jean-Louis

    2010-01-01

    We have developed an electrochemical immunosensor for the detection of ultratrace amounts of aflatoxin M(1) (AFM(1)) in food products. The sensor was based on a competitive immunoassay using horseradish peroxidase (HRP) as a tag. Magnetic nanoparticles coated with antibody (anti-AFM(1)) were used to separate the bound and unbound fractions. The samples containing AFM(1) were incubated with a fixed amount of antibody and tracer [AFM(1) linked to HRP (conjugate)] until the system reached equilibrium. Competition occurs between the antigen (AFM(1)) and the conjugate for the antibody. Then, the mixture was deposited on the surface of screen-printed carbon electrodes, and the mediator [5-methylphenazinium methyl sulphate (MPMS)] was added. The enzymatic response was measured amperometrically. A standard range (0, 0.005, 0.01, 0.025, 0.05, 0.1, 0.25, 0.3, 0.4 and 0.5 ppb) of AFM(1)-contaminated milk from the ELISA kit was used to obtain a standard curve for AFM(1). To test the detection sensitivity of our sensor, samples of commercial milk were supplemented at 0.01, 0.025, 0.05 or 0.1 ppb with AFM(1). Our immunosensor has a low detection limit (0.01 ppb), which is under the recommended level of AFM(1) [0.05 μg L-1 (ppb)], and has good reproducibility. PMID:22163418

  5. Electrochemical Urea Biosensor Based on Sol-gel Derived Nanostructured Cerium Oxide

    NASA Astrophysics Data System (ADS)

    Ansari, Anees A.; Azahar, Md; Malhotra, B. D.

    2012-04-01

    Urease (Urs) and glutamate dehydrogenase (GLDH) have been co-immobilized onto a nanostructured-cerium oxide (Nano-CeO2) film deposited onto a indium-tin-oxide (ITO) coated glass substrate by dip-coating via sol-gel process for urea detection. This nanostructured film has characterized using X-ray diffraction (XRD), Fourier transform infrared (FTIR), Scanning electron microscope (SEM) and electrochemical techniques, respectively. The particle size of the Nano-CeO2 film has been found to be 23 nm. Electrochemcial response (CV) studies show that Ur-GLDH/Nano-CeO2/ITO bioelectrode is found to be sensitive in the 10-80 mg/dL urea concentration range and can detect urea concentration upto 0.1 mg/dL level. The value of Michaelis-Menten constant (Km) estimated using Lineweaver-Burke plot found as 6.09 mg/dL indicates enhancement in the affinity and/or activity of enzyme attached to their nanobiocomposite. This bioelectrode retained 95% of enzyme activity after 6 months at 4°C.

  6. Poly(o-anisidine) films on mild steel: electrochemical synthesis and biosensor application

    NASA Astrophysics Data System (ADS)

    Patil, Dewyani; Gaikwad, A. B.; Patil, Pradip

    2007-04-01

    Poly(o-anisidine) (POA) films were synthesized on mild steel from an aqueous oxalic acid solution by electrochemical polymerization of o-anisidine using cyclic voltammetry. These films were characterized by cyclic voltammetry, UV-visible absorption spectroscopy, Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). The enzyme glucose oxidase (GOx) was entrapped into the POA film by a physical adsorption method. The resulting POA-GOx films were characterized by UV-visible absorption spectroscopy, FTIR and SEM. The amperometric response of the POA-GOx films was measured as a function of glucose concentration in phosphate buffer solution (pH 7.3). The POA-GOx films exhibited a fast amperometric response (1-5 s) and a linear response in the range of 2-20 mM glucose. The maximum current density and Michaelis-Menten constant of POA/GOx films were found to be ~406 µA cm-2 and 1.03 mM, respectively. The shelf stability, operational stability and thermal stability of these films were also investigated.

  7. Electrochemical Biosensor for Rapid and Sensitive Detection of Magnetically Extracted Bacterial Pathogens

    PubMed Central

    Setterington, Emma B.; Alocilja, Evangelyn C.

    2012-01-01

    Biological defense and security applications demand rapid, sensitive detection of bacterial pathogens. This work presents a novel qualitative electrochemical detection technique which is applied to two representative bacterial pathogens, Bacillus cereus (as a surrogate for B. anthracis) and Escherichia coli O157:H7, resulting in detection limits of 40 CFU/mL and 6 CFU/mL, respectively, from pure culture. Cyclic voltammetry is combined with immunomagnetic separation in a rapid method requiring approximately 1 h for presumptive positive/negative results. An immunofunctionalized magnetic/polyaniline core/shell nano-particle (c/sNP) is employed to extract target cells from the sample solution and magnetically position them on a screen-printed carbon electrode (SPCE) sensor. The presence of target cells significantly inhibits current flow between the electrically active c/sNPs and SPCE. This method has the potential to be adapted for a wide variety of target organisms and sample matrices, and to become a fully portable system for routine monitoring or emergency detection of bacterial pathogens. PMID:25585629

  8. Functional Polymers in Protein Detection Platforms: Optical, Electrochemical, Electrical, Mass-Sensitive, and Magnetic Biosensors

    PubMed Central

    Hahm, Jong-in

    2011-01-01

    The rapidly growing field of proteomics and related applied sectors in the life sciences demands convenient methodologies for detecting and measuring the levels of specific proteins as well as for screening and analyzing for interacting protein systems. Materials utilized for such protein detection and measurement platforms should meet particular specifications which include ease-of-mass manufacture, biological stability, chemical functionality, cost effectiveness, and portability. Polymers can satisfy many of these requirements and are often considered as choice materials in various biological detection platforms. Therefore, tremendous research efforts have been made for developing new polymers both in macroscopic and nanoscopic length scales as well as applying existing polymeric materials for protein measurements. In this review article, both conventional and alternative techniques for protein detection are overviewed while focusing on the use of various polymeric materials in different protein sensing technologies. Among many available detection mechanisms, most common approaches such as optical, electrochemical, electrical, mass-sensitive, and magnetic methods are comprehensively discussed in this article. Desired properties of polymers exploited for each type of protein detection approach are summarized. Current challenges associated with the application of polymeric materials are examined in each protein detection category. Difficulties facing both quantitative and qualitative protein measurements are also identified. The latest efforts on the development and evaluation of nanoscale polymeric systems for improved protein detection are also discussed from the standpoint of quantitative and qualitative measurements. Finally, future research directions towards further advancements in the field are considered. PMID:21691441

  9. Scanning electrochemical microscopy of genomic DNA microarrays--study of adsorption and subsequent interactions.

    PubMed

    Roberts, William S; Davis, Frank; Higson, Séamus P J

    2009-07-01

    The adsorption of genomic DNA and subsequent interactions between adsorbed and solvated DNA have been studied using scanning electrochemical microscopy (SECM). Microarrays of polyethylenimine (PEI) films could be deposited on screen-printed carbon substrates using the SECM. Single stranded herring DNA was electrostatically adsorbed at the surface of the polyethylenimine. The further adsorption of complementary single stranded DNA on the surface was observed to give rise to substantial decreases in interfacial impedance at the surface as measured by increases of tip current of the order of 1-2 nA (6%). Conversely adsorption of DNA from alternate species, i.e. salmon ssDNA on herring ssDNA, yielded much smaller changes in tip current of 0.2 nA. The significance of this work is that the approach opens up the possibility for direct label-free electrochemical interrogation of DNA microarrays as an alternative to other existing optical techniques. PMID:19562194

  10. Electrochemical sensor for multiplex screening of genetically modified DNA: identification of biotech crops by logic-based biomolecular analysis.

    PubMed

    Liao, Wei-Ching; Chuang, Min-Chieh; Ho, Ja-An Annie

    2013-12-15

    Genetically modified (GM) technique, one of the modern biomolecular engineering technologies, has been deemed as profitable strategy to fight against global starvation. Yet rapid and reliable analytical method is deficient to evaluate the quality and potential risk of such resulting GM products. We herein present a biomolecular analytical system constructed with distinct biochemical activities to expedite the computational detection of genetically modified organisms (GMOs). The computational mechanism provides an alternative to the complex procedures commonly involved in the screening of GMOs. Given that the bioanalytical system is capable of processing promoter, coding and species genes, affirmative interpretations succeed to identify specified GM event in terms of both electrochemical and optical fashions. The biomolecular computational assay exhibits detection capability of genetically modified DNA below sub-nanomolar level and is found interference-free by abundant coexistence of non-GM DNA. This bioanalytical system, furthermore, sophisticates in array fashion operating multiplex screening against variable GM events. Such a biomolecular computational assay and biosensor holds great promise for rapid, cost-effective, and high-fidelity screening of GMO. PMID:23893064

  11. Automated microfluidically controlled electrochemical biosensor for the rapid and highly sensitive detection of Francisella tularensis.

    PubMed

    Dulay, Samuel B; Gransee, Rainer; Julich, Sandra; Tomaso, Herbert; O'Sullivan, Ciara K

    2014-09-15

    Tularemia is a highly infectious zoonotic disease caused by a Gram-negative coccoid rod bacterium, Francisella tularensis. Tularemia is considered as a life-threatening potential biological warfare agent due to its high virulence, transmission, mortality and simplicity of cultivation. In the work reported here, different electrochemical immunosensor formats for the detection of whole F. tularensis bacteria were developed and their performance compared. An anti-Francisella antibody (FB11) was used for the detection that recognises the lipopolysaccharide found in the outer membrane of the bacteria. In the first approach, gold-supported self-assembled monolayers of a carboxyl terminated bipodal alkanethiol were used to covalently cross-link with the FB11 antibody. In an alternative second approach F(ab) fragments of the FB11 antibody were generated and directly chemisorbed onto the gold electrode surface. The second approach resulted in an increased capture efficiency and higher sensitivity. Detection limits of 4.5 ng/mL for the lipopolysaccharide antigen and 31 bacteria/mL for the F. tularensis bacteria were achieved. Having demonstrated the functionality of the immunosensor, an electrode array was functionalised with the antibody fragment and integrated with microfluidics and housed in a tester set-up that facilitated complete automation of the assay. The only end-user intervention is sample addition, requiring less than one-minute hands-on time. The use of the automated microfluidic set-up not only required much lower reagent volumes but also the required incubation time was considerably reduced and a notable increase of 3-fold in assay sensitivity was achieved with a total assay time from sample addition to read-out of less than 20 min. PMID:24747573

  12. Biosensor based on ultrasmall MoS2 nanoparticles for electrochemical detection of H2O2 released by cells at the nanomolar level.

    PubMed

    Wang, Tanyuan; Zhu, Haichuan; Zhuo, Junqiao; Zhu, Zhiwei; Papakonstantinou, Pagona; Lubarsky, Gennady; Lin, Jian; Li, Meixian

    2013-11-01

    Monodispersed surfactant-free MoS2 nanoparticles with sizes of less than 2 nm were prepared from bulk MoS2 by simple ultrasonication and gradient centrifugation. The ultrasmall MoS2 nanoparticles expose a large fraction of edge sites, along with their high surface area, which lead to attractive electrocatalytic activity for reduction of H2O2. An extremely sensitive H2O2 biosensor based on MoS2 nanoparticles with a real determination limit as low as 2.5 nM and wide linear range of 5 orders of magnitude was constructed. On the basis of this biosensor, the trace amount of H2O2 released from Raw 264.7 cells was successfully recorded, and an efficient glucose biosensor was also fabricated. Since H2O2 is a byproduct of many oxidative biological reactions, this work serves as a pathway for the application of MoS2 in the fields of electrochemical sensing and bioanalysis. PMID:24067077

  13. Polymerase chain reaction-free detection of hepatitis B virus DNA using a nanostructured impedance biosensor.

    PubMed

    Chen, Chun-Cheng; Lai, Zi-Lun; Wang, Gou-Jen; Wu, Chun-Ying

    2016-03-15

    A polymerase chain reaction (PCR)-free technique for the effective detection of genomic length hepatitis B virus (HBV) DNA is described in this study. The honeycomb-like barrier layer of an anodic aluminum oxide (AAO) film having a uniform nanohemisphere array was used as the substrate of the sensing electrode. A 30-nm gold film was sputtered onto the AAO barrier layer surface as the electrode, followed by electrochemical deposition of gold nanoparticles (GNPs) on the hemisphere surface. A specially designed single-strand 96-mer gene fragment of the target genomic DNA of HBV based on the genome sequences of HBV was immobilized on the nanostructured electrode as the capture probe. Target HBV DNA obtained from clinical samples was hybridized to the sensing probes. Detection results illustrate two dynamic linear ranges, 10(2)-10(3) and 10(3)-10(5.1) copies/mL, having R(2) values of 0.801 and 0.996 could be obtained, respectively. The detection limit of the proposed sending scheme was measured to be 111 copies/mL. The total of 45 target samples, including 20 samples with HBV concentration being lower than 10(2) copies/mL and 25 samples with HBV concentration being in the range of 10(3)-10(5.1) copies/mL, were used for real test. The concentration of these 45 HBV DNA samples was measured by the COBAS Ampliprep system. Comparing the measured results of the COBAS Ampliprep and our system, it was illustrated that the HBV DNA concentrations measured by the proposed method in this study had a high linear correlation with the COBAS Ampliprep, having R(2) values of 0.983. The proposed sensing scheme is highly feasible for future clinical applications. PMID:26479905

  14. DNA-functionalized silicon nitride nanopores for sequence-specific recognition of DNA biosensor

    NASA Astrophysics Data System (ADS)

    Tan, Shengwei; Wang, Lei; Yu, Jingjing; Hou, Chuanrong; Jiang, Rui; Li, Yanping; Liu, Quanjun

    2015-05-01

    Nanopores have been proven to be novel and versatile single-molecule sensors for individual unlabeled biopolymer detection and characterization. In the present study, a relatively large silicon nitride (Si3N4) nanopore with a diameter of approximately 60 nm was fabricated successfully using a focused Ga ion beam (FIB). We demonstrated a simple ex situ silanization procedure to control the size and functionality of solid-state nanopores. The presented results show that by varying the silanization time, it is possible to adjust the efficiency of probe molecule attachment, thus shrinking the pore to the chosen size, while introducing selective sensing probes. The functionalization of nanopores was verified by analysis of field-emission scanning electron microscopy (FESEM), energy-dispersive X-ray spectroscopy (EDS), and electrical measurements. Based on this study, we envision that the functionalized silicon nitride nanopores with the DNA probe might provide a biosensing platform for the detection and discrimination of a short single-stranded DNA oligomer of unknown sequences in the future.

  15. A localized surface plasmon resonance DNA biosensor based on gold nanospheres coated on the tip of the fiber

    NASA Astrophysics Data System (ADS)

    Jia, Shuo; Bian, Chao; Tong, Jian-hua; Sun, Ji-zhou; Xia, Shan-hong

    2016-03-01

    A localized surface plasmon resonance (LSPR) biosensor was prepared with gold nanospheres (AuNSs) coated on the tip face of the optical silica fiber. AuNSs with the sizes of 20 nm and 80 nm were used. The sensitivities of AuNS20 nm and AuNS80 nm modified sensors to bulk refractive index (RI) variation are 82.86 nm/RIU and 218.98 nm/RIU, respectively. The AuNS80 nm modified sensor was used for the detection of 40 bases DNA hybridization and the limit of detection is 50 nmol/L, where the 40-bases DNA probe was covalently linked with AuNS80 nm. The complementary DNA sequence in tris-acetate-EDTA (TAE) buffer solution was detected as the target DNA. This fiber sensor has the advantages of small sample consumption, easy fabrication and high sensitivity.

  16. Electrochemical properties of interstrand cross-linked DNA duplexes labeled with Nile blue.

    PubMed

    Mie, Yasuhiro; Kowata, Keiko; Kojima, Naoshi; Komatsu, Yasuo

    2012-12-11

    DNA molecules have attracted considerable attention as functional materials in various fields such as electrochemical sensors with redox-labeled DNA. However, the recently developed interstrand cross-link (ICL) technique for double-stranded DNA can adequately modify the electronic properties inside the duplex. Hence, the electrochemical investigation of ICL-DNA helps us to understand the electron transfer of redox-labeled DNA at an electrode surface, which would develop useful sensors. In this study, the first insight into this matter is presented. We prepared 17-mer DNA duplexes incorporating Nile blue (NB-DNA) at one end as a redox marker and a disulfide tether at the other end for immobilization onto an electrode. The duplexes were covalently cross-linked by bifunctional cross-linkers that utilize either a propyl or naphthalene residue to replace a base pair. Their electrochemical responses at the electrode surface were compared to evaluate the effect of the ICL on the electron-transfer reactions of the redox-labeled DNA duplexes. A direct transfer of electrons between NB and the electrode was observed for a standard DNA, as previously reported, whereas interstrand cross-linked DNA (CL-DNA) strands showed a decrease in the direct electron-transfer pathway. This is expected to result from constraining the elastic bending/flexibility of the duplex caused by the covalent cross-links. Interestingly, the CL-DNA incorporating naphthalene residues exhibited additional voltammetric peaks derived from DNA-mediated electron transfer (through base π stacking), which was not observed in the mismatched CL-DNA. The present results indicate that the ICL significantly affects electron transfer in the redox-labeled DNA at the electrode and can be an important determinant for electrochemical signaling in addition to its role in stabilizing the duplex structure. PMID:23153070

  17. Lipoxygenase-modified Ru-bpy/graphene oxide: Electrochemical biosensor for on-farm monitoring of non-esterified fatty acid.

    PubMed

    Veerapandian, Murugan; Hunter, Robert; Neethirajan, Suresh

    2016-04-15

    Elevated concentrations of non-esterified fatty acids (NEFA) in biological fluids are recognized as critical biomarkers for early diagnosis of dairy cow metabolic diseases. Herein, a cost-effective, electrochemically active, and bio-friendly sensor element based on ruthenium bipyridyl complex-modified graphene oxide nanosheets ([Ru(bpy)3](2+)-GO) is proposed as a biosensor platform for NEFA detection. Electrochemical analysis demonstrates that the [Ru(bpy)3](2+)-GO electrodes exhibit superior and durable redox properties compared to the pristine carbon and GO electrodes. Target specificity is accomplished through immobilization of the enzyme, lipoxygenase, which catalyzes the production of redox active species from NEFA. Lipoxygenases retain their catalytic ability upon immobilization and exhibit changes to amperometric signals upon interaction with various concentrations of standard NEFA and serum samples. Our study demonstrates that the [Ru(bpy)3](2+)-GO electrode has the potential to serve as a biosensor platform for developing a field deployable, rapid, and user-friendly detection tool for on-farm monitoring of dairy cow metabolic diseases. PMID:26623509

  18. Micro-patterning of ionic reservoirs within a double bilayer lipid membrane to fabricate a 2D array of ion-channel switch based electrochemical biosensors

    SciTech Connect

    Sansinena, J. M.; Yee, C. K.; Sapuri, A.; Swanson, Basil I.; Redondo, A.; Parikh, A. N.

    2004-01-01

    We present a simple approach for the design of ionic reservoir arrays within a double phospholipid bilayer to ultimately develop a 2D array of ion-channel switch based electrochemical biosensors. As a first step, a primary bilayer lipid membrane is deposited onto an array of electrodes patterned onto a substrate surface. Subsequently, an array of microvoids is created within the bilayer by a wet photolithographic patterning of phospholipid bilayers using a deep UV light source and a quartz/chrome photomask. To ensure registry, the photomask used to pattern bilayers is designed to match up the microvoids within the primary bilayer with the array of electrodes on the substrate surface. The deposition of a secondary bilayer lipid membrane onto the primary bilayer that spans across the patterned microvoids leads to the formation of the array of ionic reservoirs within the double phospholipid bilayer. This is accomplished using giant unilamellar vesicles and by exploiting membrane electrostatics. The use of ion-channels incorporated into the secondary bilayer that covers the individual ionic reservoirs allows the construction of a 2D array of ion-channel switch based electrochemical biosensors that are able to recognize different target-agents simultaneously.

  19. Novel electrochemical dual-aptamer-based sandwich biosensor using molybdenum disulfide/carbon aerogel composites and Au nanoparticles for signal amplification.

    PubMed

    Fang, Lin-Xia; Huang, Ke-Jing; Liu, Yang

    2015-09-15

    A new electrochemical aptamer biosensor for the platelet-derived growth factor BB (PDGF-BB) detection has been developed based on the signal amplification of MoS2/carbon aerogel composites (MoS2/CA) and sandwich assay. A facile hydrothermal route assisted by L-cysteine was applied to synthesize CA incorporated flower-like MoS2 with the large surface active sites and good conductivity. The electrochemical aptasensor was constructed by sandwiching the PDGF-BB between a glassy carbon electrode modified with thiol-terminated PDGF-BB aptamer-1 (Apt1)/gold nanoparticles (AuNPs)/MoS2/CA and the AuNPs with thiol-terminated PDGF-BB aptamer-2 (Apt2) and 6-ferrocenyl hexanethiol (Fc). Fc-AuNPs-Apt2 acted as tracer and AuNPs/MoS2/CA were utilized as the biosensor platform to immobilize a large amount of capture aptamers, owing to their layered structure and high surface-to-volume ratio. Based on the sandwich format, a dual signal amplification strategy had been successfully developed with a wide linear response in the range of 0.001-10nM and a limit of detection of 0.3 pM. The developed assay demonstrated good selectivity and high sensitivity, indicating potential applications in bioanalysis and biomedicine. PMID:25909336

  20. Glycan and lectin biosensors

    PubMed Central

    Belický, Štefan; Katrlík, Jaroslav

    2016-01-01

    A short description about the importance of glycan biorecognition in physiological (blood cell type) and pathological processes (infections by human and avian influenza viruses) is provided in this review. Glycans are described as much better information storage media, compared to proteins or DNA, due to the extensive variability of glycan structures. Techniques able to detect an exact glycan structure are briefly discussed with the main focus on the application of lectins (glycan-recognising proteins) in the specific analysis of glycans still attached to proteins or cells/viruses. Optical, electrochemical, piezoelectric and micromechanical biosensors with immobilised lectins or glycans able to detect a wide range of analytes including whole cells/viruses are also discussed. PMID:27365034

  1. Glycan and lectin biosensors.

    PubMed

    Belický, Štefan; Katrlík, Jaroslav; Tkáč, Ján

    2016-06-30

    A short description about the importance of glycan biorecognition in physiological (blood cell type) and pathological processes (infections by human and avian influenza viruses) is provided in this review. Glycans are described as much better information storage media, compared to proteins or DNA, due to the extensive variability of glycan structures. Techniques able to detect an exact glycan structure are briefly discussed with the main focus on the application of lectins (glycan-recognising proteins) in the specific analysis of glycans still attached to proteins or cells/viruses. Optical, electrochemical, piezoelectric and micromechanical biosensors with immobilised lectins or glycans able to detect a wide range of analytes including whole cells/viruses are also discussed. PMID:27365034

  2. A label-free fluorescent molecular beacon based on DNA-Ag nanoclusters for the construction of versatile Biosensors.

    PubMed

    Cao, Qiao; Teng, Ye; Yang, Xuan; Wang, Jin; Wang, Erkang

    2015-12-15

    In this paper, we developed a simple, low-cost and sensitive DNA sequences detection biosensor based on a label-free molecular beacon (MB) whose DNA hairpin structure terminal has a guanine-rich sequence that can enhance fluorescence of silver nanoclusters (Ag NCs). Without hybridization between hairpin probe and target DNA, the Ag NCs presented bright fluorescence for the proximity of guanine-rich sequences (GRSs). After binding with target DNA, the hairpin shape was destroyed which results in a decrease of the Ag NCs fluorescence intensity. With this biosensor, we detected three disease-related genes that were the human immunodeficiency virus (HIV) gene, hepatitis B virus (HBV) gene and human T-lymphotropic virus type I (HTLV-I) gene. The detection limits based on S/N of 3 were 4.4 nM, 6.8 nM and 8.5 nM for HIV gene, HBV gene and HTLV-I gene, respectively. Our sensor was also of high selectivity and could distinguish even one nucleotide mismatched target. PMID:26159151

  3. Micro pH Sensors and Biosensors Based on Electrochemical Field Effect Transistors

    NASA Astrophysics Data System (ADS)

    Sasano, Junji; Niwa, Daisuke; Osaka, Tetsuya

    A study on ion-sensing using field effect transistor (FET) was begun by Bergveld in the 1970s [1-3]. The ion-sensitive (IS) FET is now widely used as a miniaturized pH sensor, commercialized by some companies. First, the principle and structure of the ISFET are introduced in this section. A basic design of ISFET is shown in Fig. 10.1 a. ISFET has silicon substrate with field-effect structures such as electrolyte/IS layer/(insulator)/semiconductor structures; the space charge region in the semiconductor is modulated depending on the gate voltage (V g), same as a typical metal-oxide-semiconductor (MOS) FET. A typical bias V g versus drain-source current (I ds) characteristic of the device that has silicon nitride/silicon dioxide/silicon is shown in Fig. 10.1 b. This characteristic is quite similar to the MOSFET. A prominent difference between ISFET and MOSFET is that the gate voltage for the operation of the device is applied by an electrochemical reference electrode through the electrolyte in contact with the gate insulator. The threshold voltage (V th) could shift according to the value of the pH of the solution. In the MOSFET, the V th would shift depending on the change in the space charge region in the MOS capacitor structure by the application of V g. On the other hand, the V th in ISFET would shift according to the change in the surface potential in the electrolyte/IS layer interface. Therefore, the IS layers and their interfaces in ISFET play an important role in the performance of pH responsibility. It is well-known that the silicon nitride surface shows a good pH response in solution. The silicon nitride layer is often formed by plasma-enhanced chemical vapor deposition (PECVD), which is generally formed at the thickness of 100-500 nm. The V g vs. I ds, characteristics of the silicon nitride-based ISFET indicate a good pH responsibility of 58 mV/decade that shows Nernstian response (Fig. 10.1 c). The shift of the V th depends on the changes of surface

  4. Synthesis and characterization of reduced graphene oxide supported gold nanoparticles-poly(pyrrole-co-pyrrolepropylic acid) nanocomposite-based electrochemical biosensor.

    PubMed

    Puri, Nidhi; Niazi, Asad; Srivastava, Avanish K; Rajesh

    2014-10-01

    A conducting poly(pyrrole-co-pyrrolepropylic acid) copolymer nanocomposite film (AuNP-PPy-PPa) incorporating gold nanoparticles (AuNP) was electrochemically grown using a single step procedure over electrochemically reduced graphene oxide (RGO) flakes deposited on a silane-modified indium-tin-oxide (ITO) glass plate. The RGO support base provided excellent mechanical and chemical stability to the polymer nanocomposite matrix. The porous nanostructure of AuNP-PPy-PPa/RGO provided a huge accessible area to disperse AuNP, and it avoided metallic agglomeration within the polymer matrix. The AuNP-PPy-PPa/RGO was characterized by high-resolution transmission electron microscopy (HRTEM), contact angle measurements, Fourier transform infrared spectroscopy (FTIR), and electrochemical techniques. The pendant carboxyl group of AuNP-PPy-PPa/RGO was covalently bonded with myoglobin protein antibody, Ab-Mb, for the construction of a bioelectrode. Electrochemical impedance spectroscopy technique was used for the characterization of the bioelectrode and as an impedimetric biosensor for the detection of human cardiac biomarker, Ag-cMb. The bioelectrode exhibited a linear impedimetric response to Ag-cMb in the range of 10 ng mL(-1) to 1 μg mL(-1), in phosphate-buffered solution (PBS) (pH 7.4, 0.1 M KCl) with a sensitivity of 92.13 Ω cm(2) per decade. PMID:24928550

  5. Highly sensitive electrochemical biosensor for bisphenol A detection based on a diazonium-functionalized boron-doped diamond electrode modified with a multi-walled carbon nanotube-tyrosinase hybrid film.

    PubMed

    Zehani, Nedjla; Fortgang, Philippe; Saddek Lachgar, Mohamed; Baraket, Abdoullatif; Arab, Madjid; Dzyadevych, Sergei V; Kherrat, Rochdi; Jaffrezic-Renault, Nicole

    2015-12-15

    A highly sensitive electrochemical biosensor for the detection of Bisphenol A (BPA) in water has been developed by immobilizing tyrosinase onto a diazonium-functionalized boron doped diamond electrode (BDD) modified with multi-walled carbon nanotubes (MWCNTs). The fabricated biosensor exhibits excellent electroactivity towards o-quinone, a product of this enzymatic reaction of BPA oxidation catalyzed by tyrosinase. The developed BPA biosensor displays a large linear range from 0.01 nM to 100 nM, with a detection limit (LOD) of 10 pM. The feasibility of the proposed biosensor has been demonstrated on BPA spiked water river samples. Therefore, it could be a promising and reliable analytical tool for on-site monitoring of BPA in waste water. PMID:26232678

  6. An electrochemical DNA sensor based on polyaniline/graphene: high sensitivity to DNA sequences in a wide range.

    PubMed

    Zheng, Qing; Wu, Hao; Shen, Zongxu; Gao, Wenyu; Yu, Yu; Ma, Yuehui; Guang, Weijun; Guo, Quangui; Yan, Rui; Wang, Junzhong; Ding, Kejian

    2015-10-01

    A label-free electrochemical DNA sensor was fabricated by deposition of polyaniline and pristine graphene nanosheet (P/G(ratios)) composites in different mass ratios, DNA probe and bovine serum albumin (BSA) layer by layer on the surface of a glassy carbon electrode (GCE). Electrochemical impedance spectroscopy (EIS) was employed to monitor every step of fabrication of P/G(ratio)-based DNA sensors and to evaluate the detection results in terms of the hybridization of complementary DNA, mutant DNA and non-complementary DNA. The results illustrate that the P/G(ratio)-based DNA sensor could highly efficiently detect complementary DNA from 0.01 pm to 1 μm and discriminate single-nucleotide polymorphisms (SNPs). In the process of detection, double-stranded DNA (dsDNA), resulting from hybridization of a DNA probe, escaping from or remaining on the sensor surface, was monitored by changing the ratio of polyaniline (PANI) to graphene, which was decided by the competition between the electrostatic interaction and Brownian motion. PMID:26309910

  7. Development of a Mass Sensitive Quartz Crystal Microbalance (QCM)-Based DNA Biosensor Using a 50 MHz Electronic Oscillator Circuit

    PubMed Central

    García-Martinez, Gonzalo; Bustabad, Enrique Alonso; Perrot, Hubert; Gabrielli, Claude; Bucur, Bogdan; Lazerges, Mathieu; Rose, Daniel; Rodriguez-Pardo, Loreto; Fariña, Jose; Compère, Chantal; Vives, Antonio Arnau

    2011-01-01

    This work deals with the design of a high sensitivity DNA sequence detector using a 50 MHz quartz crystal microbalance (QCM) electronic oscillator circuit. The oscillator circuitry is based on Miller topology, which is able to work in damping media. Calibration and experimental study of frequency noise are carried out, finding that the designed sensor has a resolution of 7.1 ng/cm2 in dynamic conditions (with circulation of liquid). Then the oscillator is proved as DNA biosensor. Results show that the system is able to detect the presence of complementary target DNAs in a solution with high selectivity and sensitivity. DNA target concentrations higher of 50 ng/mL can be detected. PMID:22164037

  8. Development of a mass sensitive quartz crystal microbalance (QCM)-based DNA biosensor using a 50 MHz electronic oscillator circuit.

    PubMed

    García-Martinez, Gonzalo; Bustabad, Enrique Alonso; Perrot, Hubert; Gabrielli, Claude; Bucur, Bogdan; Lazerges, Mathieu; Rose, Daniel; Rodriguez-Pardo, Loreto; Fariña, Jose; Compère, Chantal; Vives, Antonio Arnau

    2011-01-01

    This work deals with the design of a high sensitivity DNA sequence detector using a 50 MHz quartz crystal microbalance (QCM) electronic oscillator circuit. The oscillator circuitry is based on Miller topology, which is able to work in damping media. Calibration and experimental study of frequency noise are carried out, finding that the designed sensor has a resolution of 7.1 ng/cm(2) in dynamic conditions (with circulation of liquid). Then the oscillator is proved as DNA biosensor. Results show that the system is able to detect the presence of complementary target DNAs in a solution with high selectivity and sensitivity. DNA target concentrations higher of 50 ng/mL can be detected. PMID:22164037

  9. A hairpin DNA aptamer coupled with groove binders as a smart switch for a field-effect transistor biosensor.

    PubMed

    Goda, Tatsuro; Miyahara, Yuji

    2012-02-15

    We report here that a hairpin-structured DNA that possesses an anti-ATP aptamer sequence successfully detected target ATP or adenosine in a temperature-dependent manner by nanoscale intramolecular displacement on the surface of a gold electrode as an extended gate of a field-effect transistor (FET). The structural switching of the hairpin aptamer from closed loop to open-loop conformations was accompanied by the release of the preloaded DNA binder (DAPI) from the stem part of the hairpin aptamer into the solution phase. The loss of intrinsic positive charges of DAPI (2+) from the diffusion layer at the gate/solution nano-interface as a result of target capturing was responsible for generating a specific signal by the field-effect. We emphasize a new aspect of the structured DNA aptamer in combination with FET: the DAPI-loaded hairpin aptamer successfully detected even uncharged adenosine, which remains a major challenge for FET-based biosensors. Given the simplicity in design of the primary and secondary structures of oligonucleotide aptamers, it is easy to apply this technology to a wide variety of bio-analytes, irrespective of their electric charges. In view of these advantages, our findings may offer a new trend in the design of stimuli-responsive "smart" biomolecular switches for semiconductor-based biosensors. PMID:22221798

  10. Development of Advanced Electrochemical Sensors for DNA Detection at the Point of Care

    NASA Astrophysics Data System (ADS)

    Hsieh, Kuangwen

    In the post-genomic era, ever-advancing capabilities in DNA detection and analysis have become vital to the detection of infectious diseases and the diagnosis of genetic abnormalities and inheritable diseases. The benefit of such capabilities, however, has yet to reach patients outside of centralized facilities. There thus exists an increasing need to decentralize DNA detection methods and to administer such diagnostics at the "point of care." Electrochemical-based DNA sensors present a compelling approach, but have yet to deliver satisfactory sensitivity, specificity, miniaturization, and real-time monitoring capability to meet the demand of point-of-care diagnostics. Motivated by their potential and their current limitations, in this dissertation, we present a series of strategies that we have undertaken in order to address the key shortcomings of electrochemical DNA sensors and advance them toward point-of-care applications. First, we report a single-step, single reagent, label-free, isothermal electrochemical DNA sensor based on the phenomenon of enzyme catalyzed target recycling amplification. Using this technique, we achieve improved detection limit in comparison to hybridization-based sensors without amplification. We also demonstrate greater than 16-fold amplification of signal at low target concentrations. Next, we present a novel electrochemical DNA sensor that detects single-nucleotide mismatched targets with unprecedented "polarity-switching" responses. This "bipolar" sensor employs a surface-bound and redox-modified (methylene blue) DNA probe architecture, and outputs a decreased Faradaic current when hybridized to a perfectly matched (PM) target, but conversely reports an increased Faradaic current when hybridized to a single-base mismatched (SM) target. Third, we describe the microfluidic electrochemical dynamic allele specific hybridization (microE-DASH) platform for versatile and rapid detection of single-nucleotide polymorphisms. Implementing

  11. Sensitive and visual detection of sequence-specific DNA-binding protein via a gold nanoparticle-based colorimetric biosensor.

    PubMed

    Ou, Li-Juan; Jin, Pei-Yan; Chu, Xia; Jiang, Jian-Hui; Yu, Ru-Qin

    2010-07-15

    A novel exonuclease III (Exo III) protection-based colorimetric biosensing strategy was developed for rapid, sensitive, and visual detection of sequence-specific DNA-binding proteins. This strategy relied on the protection of DNA-cross-linked gold nanoparticle (AuNP) aggregates from Exo III-mediated digestion by specific interactions of target proteins with their binding sequences. Interestingly, we disclosed a new finding that binding of target proteins to their binding sequences in the aggregated AuNP network rendered a stable and long-period protection of DNA. Unlike conventional fluorescence assays merely based on temporal protection of DNA from Exo III digestion, the stable protection afforded a static color transition indicator for DNA-protein interactions with no time-dependent monitoring required in the assay. Therefore, it furnished the developed strategy with improved technical robustness and operational convenience. Furthermore, we introduced thioctic acid as a stable anchor for tethering DNA on AuNPs. This DNA-tethering protocol circumvented the interferences from thiol compounds in common enzymatic systems. The Exo III protection-based colorimetric biosensor was demonstrated using a model target of TATA binding protein, a key transcriptional factor involving in various transcriptional regulatory networks. The results revealed that the method allowed a specific, simple, and quantitative assay of the target protein with a linear response range from 0 to 120 nM and a detection limit of 10 nM. PMID:20565105

  12. Highly sensitive silicon nanowire biosensor with novel liquid gate control for detection of specific single-stranded DNA molecules.

    PubMed

    Adam, Tijjani; Hashim, U

    2015-05-15

    The study demonstrates the development of a liquid-based gate-control silicon nanowire biosensor for detection of specific single-stranded DNA (ssDNA) molecules. The sensor was fabricated using conventional photolithography coupled with an inductively coupled plasma dry etching process. Prior to the application of DNA to the device, its linear response to pH was confirmed by serial dilution from pH 2 to pH 14. Then, the sensor surface was silanized and directly aminated with (3-aminopropyl) triethoxysilane to create a molecular binding chemistry for biofunctionalization. The resulting Si‒O‒Si‒ components were functionalized with receptor ssDNA, which interacted with the targeted ssDNA to create a field across the silicon nanowire and increase the current. The sensor shows selectivity for the target ssDNA in a linear range from target ssDNA concentrations of 100 pM to 25 nM. With its excellent detection capabilities, this sensor platform is promising for detection of specific biomarkers and other targeted proteins. PMID:25453738

  13. A novel reactive epitope-based antigen targeted by serum autoantibodies in oligoarticular and polyarticular juvenile idiopathic arthritis and development of an electrochemical biosensor.

    PubMed

    Araujo, Galber R; Fujimura, Patricia T; Vaz, Emília R; Silva, Tamiris A; Rodovalho, Vinícius R; Britto-Madurro, Ana Graci; Madurro, João M; Fonseca, João E; Silva, Carlos H M; Santos, Paula S; Mourão, Ana F; Canhão, Helena; Goulart, Luiz R; Gonçalves, João; Ueira-Vieira, Carlos

    2016-05-01

    Currently, there are no specific markers for juvenile idiopathic arthritis (JIA) diagnosis, which is based on clinical symptoms and some blood tests for diseases' exclusion. Aiming to select new epitope-based antigens (mimotopes) that could recognize circulating autoantibodies in most JIA forms, we screened a phage displayed random peptide library against IgG antibodies purified from serum of JIA patients. ELISA assay was carried out to confirm immunoreactivity of selected peptides against sera IgG antibodies from JIA patients, healthy children and patients with other autoimmune diseases. The mimotope PRF+1 fused to phage particles was able to efficiently discriminate JIA patients from controls, and for this reason was chosen to be chemically synthesized for validation in a larger sample size. The synthetic peptide was immobilized onto bioelectrodes' surface for antibody detection by electrochemical analyses through differential pulse voltammetry. The PRF+1 synthetic peptide has efficiently discriminated JIA patients from control groups (p<0.0001) with a very good accuracy (AUC>0.84; sensitivity=61%; specificity=91%). The electrochemical platform proved to be fast, low cost and effective in detecting anti-PRF+1 antibodies from JIA patients compared to healthy controls (p=0.0049). Our study describes a novel and promising epitope-based biomarker for JIA diagnosis that can become a useful tool for screening tests, which was successfully incorporated onto an electrochemical biosensor and could be promptly used in field diagnostics. PMID:26806845

  14. Sensitive electrochemical assaying of DNA methyltransferase activity based on mimic-hybridization chain reaction amplified strategy.

    PubMed

    Zhang, Linqun; Liu, Yuanjian; Li, Ying; Zhao, Yuewu; Wei, Wei; Liu, Songqin

    2016-08-24

    A mimic-hybridization chain reaction (mimic-HCR) amplified strategy was proposed for sensitive electrochemically detection of DNA methylation and methyltransferase (MTase) activity In the presence of methylated DNA, DNA-gold nanoparticles (DNA-AuNPs) were captured on the electrode by sandwich-type assembly. It then triggered mimic-HCR of two hairpin probes to produce many long double-helix chains for numerous hexaammineruthenium (III) chloride ([Ru(NH3)6](3+), RuHex) inserting. As a result, the signal for electrochemically detection of DNA MTase activity could be amplified. If DNA was non-methylated, however, the sandwich-type assembly would not form because the short double-stranded DNAs (dsDNA) on the Au electrode could be cleaved and digested by restriction endonuclease HpaII (HapII) and exonuclease III (Exo III), resulting in the signal decrement. Based on this, an electrochemical approach for detection of M.SssI MTase activity with high sensitivity was developed. The linear range for M.SssI MTase activity was from 0.05 U mL(-1) to 10 U mL(-1), with a detection limit down to 0.03 U mL(-1). Moreover, this detecting strategy held great promise as an easy-to-use and highly sensitive method for other MTase activity and inhibition detection by exchanging the corresponding DNA sequence. PMID:27496999

  15. [G3T]5/Tb(3+) based DNA biosensor with target DNA-triggered autocatalytic multi-cycle-amplification and magnetic nanoparticles assisted-background-lowered.

    PubMed

    Jiang, Hong; Zhang, Xiaojun; Wang, Guangfeng

    2015-12-15

    Due to terbium's unique photophysical properties, nucleic-acid-sensitized terbium (DNA/Tb(3+)) bioluminescent system becomes a potential candidate for the fabrication of DNA biosensors. However, the low sensitivity of DNA/Tb(3+) bioluminescent system limits its development. In this paper, a strategy combining autocatalytic multi-cycle-amplification (including exonuclease III (exo III)-aided and Zn(2+)-requiring DNAzyme-assisted target recycling amplifications) and magnetic nanoparticles assisted-background-lowering to improve the sensitivity of DNA/Tb(3+) bioluminescent system is presented for sensitive detection of target DNA (tDNA). The DNA/Tb(3+) bioluminescent system was investigated by ultraviolet-visible (UV-vis) absorption and luminescence spectra. The possible conjugation mechanism and mode of DNA with Tb(3+) were discussed. The autocatalytic multi-cycle-amplification effect was investigated by the comparison of the luminescence. The carboxylation-functionalized Fe3O4-magnetic nanoparticles (MNPs) were characterized and its role in background lowering was proved. As a result, with the designed protocol, the detection limit for the tDNA detection reached a low level to aM, which is especially exciting for the DNA/Tb(3+) bioluminescent system. In the process, due to the separation effect of MNPs, the assay solution was purified to avoid the nonspecific luminescence of DNA/Tb(3+), not only lowering the background signal greatly (about five times lower than that without the use of MNPs but also improving the reproducibility and stability. We hope that our attempt in this field will not only extend the application of DNA/Tb(3+) luminescent system in biosensing areas but also open the road to adaptation of the protocols to other related analytes. PMID:26257185

  16. Genetically Engineered Phage-Templated MnO2 Nanowires: Synthesis and Their Application in Electrochemical Glucose Biosensor Operated at Neutral pH Condition.

    PubMed

    Han, Lei; Shao, Changxu; Liang, Bo; Liu, Aihua

    2016-06-01

    To conveniently obtain one-dimensional MnO2 nanowires (NWs) with controlled structure and unique properties for electron transfer, the genetically engineered M13 phages were used as templates for precise nucleation and growth of MnO2 crystals in filamentous phage scaffolds, via the spontaneous oxidation of Mn(2+) in alkaline solution. It was found that the morphology of NWs could be tailored by the surface charge of M13 mutants. MnO2 crystals were uniformly distributed on the surface of negatively charged tetraglutamate-fused phage (M13-E4), significantly different from irregular MnO2 agglomeration on the weakly negatively charged wild-type phage and positively charged tetraarginine-fused phage. The as-synthesized M13-E4@MnO2 NWs could catalyze the electro-oxidation of H2O2 at neutral pH. To demonstrate the superiority of the electrocatalytic activity in the solution containing plenty of chloride ions at neutral pH, both glucose oxidase and as-prepared MnO2 NWs were used for fabricating the glucose biosensor. The proposed biosensor showed a wide linear range (5 μM to 2 mM glucose), a low limit of detection of 1.8 μM glucose (S/N = 3), good interassay and intra-assay reproducibility and satisfactory storage stability. Due to the superiorities of synthesis and electrochemical performance, the as-prepared MnO2 NWs are promising for applications in electrocatalysis, electrochemical sensor, and supercapacitor. PMID:27228383

  17. Real-time monitoring of superoxide accumulation and antioxidant activity in a brain slice model using an electrochemical cytochrome c biosensor

    PubMed Central

    Ganesana, Mallikarjunarao; Erlichman, Joseph S.; Andreescu, Silvana

    2012-01-01

    The overproduction of reactive oxygen species and resulting damage are central to the pathology of many diseases. The study of the temporal and spatial accumulation of reactive oxygen species has been limited due to the lack of specific probes and techniques capable of continuous measurement. We demonstrate the use of a miniaturized electrochemical cytochrome C (Cyt C) biosensor for real-time measurements and quantitative assessment of superoxide production and inactivation by natural and engineered antioxidants in acutely prepared brain slices from mice. During control conditions, superoxide radicals produced from the hippocampal region of the brain in 400 μm thick sections were well within the range of detection of the electrode. Exposure of the slices to ischemic conditions increased the superoxide production two fold and measurements from the slices were stable over a 3–4 hour period. The stilbene derivative and anion channel inhibitor, 4,4′-diisothiocyano-2,2′-disulfonic stilbene (DIDS), markedly reduced the extracellular superoxide signal under control conditions suggesting that a transmembrane flux of superoxide into the extracellular space may occur as part of normal redox signaling. The specificity of the electrode for superoxide released by cells in the hippocampus was verified by the exogenous addition of superoxide dismutase (SOD) which decreased the superoxide signal in a dose-dependent manner. Similar results were seen with the addition of the SOD-mimetic, cerium oxide nanoparticles (nanoceria) where the superoxide anion radical scavenging activity of nanoceria with an average diameter of 15 nm was equivalent to 527 U of SOD for each 1 μg/ml of nanoceria added. This study demonstrates the potential of electrochemical biosensors for studying real-time dynamics of reactive oxygen species in a biological model and the utility of these measurements in defining the relative contribution of superoxide to oxidative injury. PMID:23085519

  18. Development of a mediated whole cell-based electrochemical biosensor for joint toxicity assessment of multi-pollutants using a mixed microbial consortium.

    PubMed

    Gao, Guanyue; Qian, Jun; Fang, Deyu; Yu, Yuan; Zhi, Jinfang

    2016-06-14

    Since most risk assessment for toxicants is based on individual single-species test, the deduction of such results to ecosystem evaluation is afflicted with uncertainties. Herein, we successfully developed a p-benzoquinone mediated whole-cell electrochemical biosensor for multi-pollutants toxicological analysis by co-immobilizing mixed strains of microorganism, including Escherichia coli (gram-negative bacteria), Bacillus subtilis (gram-positive bacteria) and Saccharomyces cerevisiae (fungus). The individual and combined toxicities of heavy metal ions (Cu(2+), Cd(2+)), phenol (3,5-dichlorophenol) and pesticides (Ametryn, Acephate) were examined. The experimental results showed that the order of toxicity for individual toxicant was ranked as Cu(2+) > 3,5-dichlorophenol (DCP) > Ametryn > Cd(2+) > Acephate. Then the toxic unit (TU) model was applied to determine the nature of toxicological interaction of the toxicants which can be classified as concentration additive (IC50mix = 1TU), synergistic (IC50mix < 1TU) and antagonistic (IC50mix > 1TU) responses. The binary combination of Cu(2+) + Cd(2+), Cu(2+) + DCP, Cu(2+) + Acephate, DCP + Acephate, Acephate + Ametryn were analyzed and the three kind of joint toxicity effects (i.e. additive, synergistic and antagonistic) mentioned above were observed according to the dose-response relationship. The results indicate that the whole-cell electrochemical biosensor based on mixed microbial consortium is more reasonable to reflect the joint biotoxicity of multi-pollutants existing in real wastewater, and combined effects of toxicants is extremely necessary to be taken into account in ecological risk assessment. Thus, present study has provided a promising approach to the quality assessment of wastewater and a reliable way for early risk warning of acute biotoxicity. PMID:27181640

  19. Signal enhancement of silicon nanowire-based biosensor for detection of matrix metalloproteinase-2 using DNA-Au nanoparticle complexes.

    PubMed

    Choi, Jin-Ha; Kim, Han; Choi, Jae-Hak; Choi, Jeong-Woo; Oh, Byung-Keun

    2013-11-27

    Silicon nanowires have been used in the development of ultrasensitive biosensors or chemical sensors, which is originated in its high surface-to-volume ratio and function as field-effect transistor (FET). In this study, we developed an ultrasensitive DNA-gold (Au) nanoparticle complex-modified silicon nanowire field effect transistor (SiNW-FET) biosensor to detect matrix metalloproteinase-2 (MMP-2), which has been of particular interest as protein biomarker because of its relation to several important human diseases, through an enzymatic cleavage reaction of a specific peptide sequence (IPVSLRSG). SiNW patterns with a width of 100 nm and height of 100 nm were fabricated on a p-type silicon-on-insulator (SOI) wafer by electron-beam lithography. Next, negatively charged DNA-Au nanoparticle complexes coupled with the specific peptide (KKGGGGGG-IPVSLRSG-EEEEEE) were applied on the SiNWs to create a more sensitive system, which was then bound to aldehyde-functionalized SiNW. The enhanced negatively charged nanoparticle complexes by attached DNA were used to enhance the conductance change of the p-SiNW by MMP-2 cleavage reaction of the specific peptide. MMP-2 was successfully measured within a range of 100 fM to 10 nM, and the conductance signal of the p-type SiNW by the MMP-2 cleavage reaction was enhanced over 10-fold by using the DNA-Au nanoparticle complexes compared with using SiNW-attached negative single peptide sequences. PMID:24164583

  20. Electrochemical DNA sensor for anthrax toxin activator gene atxA-detection of PCR amplicons.

    PubMed

    Das, Ritu; Goel, Ajay K; Sharma, Mukesh K; Upadhyay, Sanjay

    2015-12-15

    We report the DNA probe functionalized electrochemical genosensor for the detection of Bacillus anthracis, specific towards the regulatory gene atxA. The DNA sensor is fabricated on electrochemically deposited gold nanoparticle on self assembled layer of (3-Mercaptopropyl) trimethoxysilane (MPTS) on GC electrode. DNA hybridization is monitored by differential pulse voltammogram (DPV). The modified GC electrode is characterized by atomic force microscopy (AFM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) method. We also quantified the DNA probe density on electrode surface by the chronocoulometric method. The detection is specific and selective for atxA gene by DNA probe on the electrode surface. No report is available for the detection of B. anthracis by using atxA an anthrax toxin activator gene. In the light of real and complex sample, we have studied the PCR amplicons of 303, 361 and 568 base pairs by using symmetric and asymmetric PCR approaches. The DNA probe of atxA gene efficiently hybridizes with different base pairs of PCR amplicons. The detection limit is found to be 1.0 pM (S/N ratio=3). The results indicate that the DNA sensor is able to detect synthetic target as well as PCR amplicons of different base pairs. PMID:26257186

  1. Insertion approach: bolstering the reproducibility of electrochemical signal amplification via DNA superstructures.

    PubMed

    Yang, Li; Zhang, Caihua; Jiang, Hong; Li, Guijuan; Wang, Jiahai; Wang, Erkang

    2014-05-20

    For more than a decade, the backfilling approach for the immobilization of DNA probes has been routinely adopted for the construction of functional interfaces; however, reliably reproducing electrochemical signal amplification by this method is a challenge. In this research, we demonstrate that the insertion approach significantly bolsters the reproducibility of electrochemical signal amplification via DNA superstructures. The combination of the backfilling approach and the DNA superstructure formation poses a big challenge to reliably reproducing electrochemical signal amplification. In order to use the detection of Hg(2+) as a prototype of this new strategy, a thymine-rich DNA probe that is specific to mercury ion was applied in this study. The presence of Hg(2+) induces the folding of the DNA probes and inhibits the formation of DNA superstructures. By using electroactive probes ([Ru(NH3)6](3+)) that are electrostatically adsorbed onto the double strands, differential pulse voltammetry (DPV) could quantitatively confirm the presence of Hg(2+). A limit of detection (LOD) and a limit of quantification (LOQ) (LOQ) as low as 0.3 and 9.5 pM, respectively, were achieved. Furthermore, excellent selectivity and real sample analysis demonstrated the promising potential of this approach in future applications. PMID:24761933

  2. Porous graphene oxide nanostructure as an excellent scaffold for label-free electrochemical biosensor: Detection of cardiac troponin I.

    PubMed

    Kazemi, Sayed Habib; Ghodsi, Elham; Abdollahi, Siamak; Nadri, Samad

    2016-12-01

    Herein, we report the fabrication of a novel label-free impedimetric biosensor employing porous graphene oxide (PrGO) nanostructures for the specific detection of cardiac troponin-I (cTnI) to establish the myocardial infarction (MI). This nano-immunosensor demonstrates an outstanding selectivity and high sensitivity towards the human-cTnI analyte. An excellent detection limit of 0.07ngmL(-1) and dynamic linear range of 0.1-10ngmL(-1) were calculated for anti-cTnI/PrGO/GC. Finally, this biosensor was employed to check the concentration of the MI biomarker in real clinical samples and the results are in good agreement with standard enzyme-linked fluorescence assay (ELFA) method. PMID:27612734

  3. Titanium Dioxide Nanoparticle-Based Interdigitated Electrodes: A Novel Current to Voltage DNA Biosensor Recognizes E. coli O157:H7

    PubMed Central

    Nadzirah, Sh.; Azizah, N.; Hashim, Uda; Gopinath, Subash C. B.; Kashif, Mohd

    2015-01-01

    Nanoparticle-mediated bio-sensing promoted the development of novel sensors in the front of medical diagnosis. In the present study, we have generated and examined the potential of titanium dioxide (TiO2) crystalline nanoparticles with aluminium interdigitated electrode biosensor to specifically detect single-stranded E.coli O157:H7 DNA. The performance of this novel DNA biosensor was measured the electrical current response using a picoammeter. The sensor surface was chemically functionalized with (3-aminopropyl) triethoxysilane (APTES) to provide contact between the organic and inorganic surfaces of a single-stranded DNA probe and TiO2 nanoparticles while maintaining the sensing system’s physical characteristics. The complement of the target DNA of E. coli O157:H7 to the carboxylate-probe DNA could be translated into electrical signals and confirmed by the increased conductivity in the current-to-voltage curves. The specificity experiments indicate that the biosensor can discriminate between the complementary sequences from the base-mismatched and the non-complementary sequences. After duplex formation, the complementary target sequence can be quantified over a wide range with a detection limit of 1.0 x 10-13M. With target DNA from the lysed E. coli O157:H7, we could attain similar sensitivity. Stability of DNA immobilized surface was calculated with the relative standard deviation (4.6%), displayed the retaining with 99% of its original response current until 6 months. This high-performance interdigitated DNA biosensor with high sensitivity, stability and non-fouling on a novel sensing platform is suitable for a wide range of biomolecular interactive analyses. PMID:26445455

  4. Titanium Dioxide Nanoparticle-Based Interdigitated Electrodes: A Novel Current to Voltage DNA Biosensor Recognizes E. coli O157:H7.

    PubMed

    Nadzirah, Sh; Azizah, N; Hashim, Uda; Gopinath, Subash C B; Kashif, Mohd

    2015-01-01

    Nanoparticle-mediated bio-sensing promoted the development of novel sensors in the front of medical diagnosis. In the present study, we have generated and examined the potential of titanium dioxide (TiO2) crystalline nanoparticles with aluminium interdigitated electrode biosensor to specifically detect single-stranded E.coli O157:H7 DNA. The performance of this novel DNA biosensor was measured the electrical current response using a picoammeter. The sensor surface was chemically functionalized with (3-aminopropyl) triethoxysilane (APTES) to provide contact between the organic and inorganic surfaces of a single-stranded DNA probe and TiO2 nanoparticles while maintaining the sensing system's physical characteristics. The complement of the target DNA of E. coli O157:H7 to the carboxylate-probe DNA could be translated into electrical signals and confirmed by the increased conductivity in the current-to-voltage curves. The specificity experiments indicate that the biosensor can discriminate between the complementary sequences from the base-mismatched and the non-complementary sequences. After duplex formation, the complementary target sequence can be quantified over a wide range with a detection limit of 1.0 x 10(-13)M. With target DNA from the lysed E. coli O157:H7, we could attain similar sensitivity. Stability of DNA immobilized surface was calculated with the relative standard deviation (4.6%), displayed the retaining with 99% of its original response current until 6 months. This high-performance interdigitated DNA biosensor with high sensitivity, stability and non-fouling on a novel sensing platform is suitable for a wide range of biomolecular interactive analyses. PMID:26445455

  5. Electrochemical biosensor based on enzyme substrate as a linker: Application for aldolase activity with pectin-thionine complex as recognization element and signal amplification probe.

    PubMed

    Wang, Xiaonan; Wang, Meiwen; Zhang, Yuanyuan; Miao, Xiaocao; Huang, Yuanyuan; Zhang, Juan; Sun, Lizhou

    2016-09-15

    A new strategy to fabricate electrochemical biosensor is reported based on the linkage of enzyme substrate, thereby an electrochemical method to detect aldolase activity is established using pectin-thionine complex (PTC) as recognization element and signal probe. The linkage effect of fructose-1,6-bisphosphate (FBP), the substrate of aldolase, can be achieved via its strong binding to magnetic nanoparticles (MNPs)/aminophenylboronic acid (APBA) and the formation of phosphoramidate bond derived from its reaction with p-phenylenediamine (PDA) on the surface of electrode. Aldolase can reversibly catalyze the substrates into the products which have no binding capacity with MNPs/APBA, resulting in the exposure of the corresponding binding sites and its subsequent recognization on signal probe. Meanwhile, signal amplification can be accomplished by using the firstly prepared PTC which can bind with MNPs/APBA, and accuracy can be strengthened through magnetic separation. With good precision and accuracy, the established sensor may be extended to other proteins with reversible catalyzed ability. PMID:27107145

  6. Carbon nanotube/polymer composite electrodes for flexible, attachable electrochemical DNA sensors.

    PubMed

    Li, Jianfeng; Lee, Eun-Cheol

    2015-09-15

    All-solution-processed, easily-made, flexible multi-walled carbon nanotube (MWCNT)/polydimethylsiloxane (PDMS)-based electrodes were fabricated and used for electrochemical DNA sensors. These electrodes could serve as a recognition layer for DNA, without any surface modification, through π-π interactions between the MWCNTs and DNA, greatly simplifying the fabrication process for DNA sensors. The electrodes were directly connected to an electrochemical analyzer in the differential pulse voltammetry (DPV) and cyclic voltammetry (CV) measurements, where methylene blue was used as a redox indicator. Since neither functional groups nor probe DNA were immobilized on the surfaces of the electrodes, the sensor can be easily regenerated by washing these electrodes with water. The limit of detection was found to be 1.3 × 10(2)pM (S/N=3), with good DNA sequence differentiation ability. Fast fabrication of a DNA sensor was also achieved by cutting and attaching the MWCNT-PDMS composite electrodes at an analyte solution-containable region. Our results pave the way for developing user-fabricated easily attached DNA sensors at low costs. PMID:25950937

  7. Smartphone-based detection of unlabeled DNA via electrochemical dissolution.

    PubMed

    Huang, Yu-Wen; Ugaz, Victor M

    2013-05-01

    We describe a novel approach that enables unlabeled biomolecules and chemical analytes to be detected using ordinary smartphone optics. Electrochemical reactivity of chromium, ordinarily considered detrimental, is harnessed here to generate a signature that can be easily seen by monitoring electrode dissolution under ordinary white-light illumination. The simplicity and robustness of this approach eliminates the need for labeling and/or pre-programming with specific receptors (e.g., oligonucleotide probes), making it feasible to greatly expand availability of a host of assays where detection complexity is a limiting constraint. PMID:23476923

  8. Electrochemical detection of the amino-substituted naphthalene compounds based on intercalative interaction with hairpin DNA by electrochemical impedance spectroscopy.

    PubMed

    Liang, Gang; Li, Tao; Li, Xiaohong; Liu, Xinhui

    2013-10-15

    The amino-substituted naphthalene compounds, such as 1,8-diaminonaphthalene (1,8-DANAP), 2,3-diaminonaphthalene (2,3-DANAP), 1,5-diaminonaphthalene (1,5-DANAP), 1-naphthylamine (1-NAP) and 2-naphthylamine (2-NAP), were investigated by electrochemical impedance spectroscopy (EIS), which was based on the interaction with hairpin DNA immobilized on the gold electrodes. Upon hairpin DNA interacting with the target chemicals, the charge transfer resistance (RCT) of the hairpin DNA films was significantly decreased and the charge transfer resistance change (ΔR(CT)) decreased in a sequence of ΔR(CT) (1,8-DANAP)>ΔR(CT) (2,3-DANAP)>ΔR(CT) (1,5-DANAP)>ΔR(CT) (1-NAP)>ΔR(CT) (2-NAP). The ΔR(CT) changes were due to the difference in the binding constant (K(SV)) of the target chemicals to DNA. In addition, the interaction mechanism was further explored using 1,8-DANAP as a model analyte by fluorescence spectra, Raman spectroscopy, differential pulse voltammetry (DPV) and EIS, correspondingly. The results demonstrated that the amino-substituted naphthalene compounds intercalated into "stem" appearing in the hairpin DNA. Moreover, the hairpin DNA sensor exhibited high sensitivity to the amino-substituted naphthalene compounds with the detection limit of nano-mole, and maintained high selectivity over other selected environmental pollutants. Finally, the DNA sensor was challenged in natural water sample with a recovery of 96-102%, which offered a platform for prospective future development of a simple, rapid, sensitive and low-cost assay for the detection of target aromatic amine pollutants. PMID:23693094

  9. Highly Selective and Sensitive Electrochemiluminescence Biosensor for p53 DNA Sequence Based on Nicking Endonuclease Assisted Target Recycling and Hyperbranched Rolling Circle Amplification.

    PubMed

    Yang, Linlin; Tao, Yingzhou; Yue, Guiyin; Li, Ruibao; Qiu, Bin; Guo, Longhua; Lin, Zhenyu; Yang, Huang-Hao

    2016-05-17

    An ultrasensitive and specific electrochemiluminescence (ECL) biosensor has been designed for the p53 DNA sequence, which is based on cascade signal amplification of nicking endonuclease assisted target recycling and hyperbranched rolling circle amplification (HRCA). First of all, biotin modified hairpin capture DNA (HP) probe was immobilized on the surface of streptavidin magnespheres paramagnetic particles (PMPs). Target DNA hybridized with the loop portion of the HP probe, therefore unfolding HP to form a double-stranded DNA (dsDNA) containing the specific nicking site of the nicking endonuclease. Then, the nicking endonuclease recognized the specific nicking site and cleaved the HP into two pieces, liberating target DNA and the complementary sequence piece for the padlock probe. The intact target DNA would initiate the next cycle of hybridization and cleavage, thereby releasing multiple complementary sequences for the padlock probes. The liberated complementary sequences hybridized with the padlock probes, subsequently inducing the HRCA reaction and generating numerous dsDNA segments. Herein, Ru(phen)3(2+) was embedded into dsDNA and worked as ECL signal reporter. The reaction products were eventually pretreated by dialysis tube with the cutoff membrane to remove the residual Ru(phen)3(2+) in the solution for the following ECL measurements. Using this cascade amplification strategy, an ultrasensitive p53 DNA sequence detection method was developed with a wide linear range from 0.05 to 100 fM and a low detection limit of 0.02 fM. Moreover, this cascade amplified ECL biosensor had specific recognition capacity for noncomplementary and single- and double-base mismatched DNA. The proposed ECL biosensor might have a great potential in biomedical research and clinic analysis. PMID:27086663

  10. Novel homogeneous label-free electrochemical aptasensor based on functional DNA hairpin for target detection.

    PubMed

    Zhang, De-Wen; Nie, Ji; Zhang, Fang-Ting; Xu, Li; Zhou, Ying-Lin; Zhang, Xin-Xiang

    2013-10-01

    We first developed a label-free and immobilization-free homogeneous electrochemical aptasensor, which combined a smart functional DNA hairpin and a designed miniaturized electrochemical device. Cocaine was chosen as a model target. The anticocaine aptamer and peroxidase-mimicking DNAzyme were integrated into one single-stranded DNA hairpin. Both aptamer and G-quadruplex were elaborately blocked by the stem region. The conformation switching induced by the affinity interaction between aptamer and cocaine released G-quadruplex part and turned on DNAzyme activity. The designed electrochemical device, constructed by a disposable micropipet tip and a reproducible carbon fiber ultramicroelectrode, was applied to the detection of homogeneous DNAzyme catalytic activity at the microliter level. The aptasensor realized the quantification of cocaine ranging from 1 to 500 μM with high specificity. The clever combination of the functional DNA hairpin and the novel device achieved an absolutely label-free electrochemical aptasensor, which showed excellent performance like low cost, easy operation, rapid detection, and high repeatability. PMID:23998357

  11. Direct electrochemical stripping detection of cystic-fibrosis-related DNA linked through cadmium sulfide quantum dots

    NASA Astrophysics Data System (ADS)

    Marin, Sergio; Merkoçi, Arben

    2009-02-01

    Electrochemical detection of a cadmium sulfide quantum dots (CdS QDs)-DNA complex connected to paramagnetic microbeads (MB) was performed without the need for chemical dissolving. The method is based on dropping 20 µl of CdS QD-DNA-MB suspension on the surface of a screen-printed electrode. It is followed by magnetic collection on the surface of the working electrode and electrochemical detection using square-wave voltammetry (SWV), giving a well-shaped and sensitive analytical signal. A cystic-fibrosis-related DNA sequence was sandwiched between the two DNA probes. One DNA probe is linked via biotin-streptavidin bonding with MB and the other one via thiol groups with the CdS QD used as tags. Nonspecific signals of DNA were minimized using a blocking agent and the results obtained were successfully employed in a model DNA sensor with an interest in future applications in the clinical field. The developed nanoparticle biosensing system may offer numerous opportunities in other fields where fast, low cost and efficient detection of small volume samples is required.

  12. An amplified electrochemical strategy using DNA-QDs dendrimer superstructure for the detection of thymine DNA glycosylase activity.

    PubMed

    Liu, Hongying; Lou, Youbing; Zhou, Fei; Zhu, Hao; Abdel-Halim, E S; Zhu, Jun-Jie

    2015-09-15

    A triple-signal amplification strategy was proposed for highly sensitive and selective detection of thymine DNA glycosylase (TDG) by coupling a dendrimer-like DNA label with the electrochemical method and quantum dots (QDs) tagging. The DNA-QDs dendrimer-like superstructure was designed by DNA hybridization and covalent assembling. Benefiting from outstanding performance of the amplification strategy, this assay showed high sensitivity, extraordinary stability, and easy operation. The limit of detection could reach 0.00003 U µL(-1) with a splendid specificity. The TDG content in different concentration of HeLa cell was also determined. This assay opens a new horizon for both qualitative and quantitative detection of TDG, holding great promise for potential application in cancer cell research and clinical diagnostics. PMID:25913445

  13. Cyclometalated iridium complex-based label-free photoelectrochemical biosensor for DNA detection by hybridization chain reaction amplification.

    PubMed

    Li, Chunxiang; Wang, Hongyang; Shen, Jing; Tang, Bo

    2015-04-21

    Photoactive material is the most crucial factor which intimately determines analytical performances of the photoelectrochemical sensor. On the basis of the high affinity of dipyrido [3,2-a:2',3'-c] phenazine (dppz) with DNA helix, a novel photoactive intercalator, [(ppy)2Ir(dppz)](+)PF6(-)(ppy = 2-phenylpyridine and dppz = dipyrido [3,2-a:2',3'-c] phenazine) was prepared and characterized by UV-vis absorption spectroscopy, fluorescence spectroscopy, and cyclic voltammetry. The photoelectrochemical properties of the as-prepared iridium(III) complex immobilized on the ITO electrode was investigated. Either cathodic or anodic photocurrent generation can be observed when triethanolamine (TEOA) or dissolved O2 is used as a sacrificial electron donor/acceptor, respectively. The probable photocurrent-generation mechanisms are speculated. A highly sensitive iridium(III) complex-based photoelectrochemical sensor was proposed for DNA detection via hybridization chain reaction (HCR) signal amplification. Under optimal conditions, the biosensor was found to be linearly proportional to the logarithm of target DNA concentration in the range from 0.025 to 100 pmol L(-1) with a detection limit of 9.0 fmol L(-1) (3σ). Moreover, the proposed sensor displayed high selectivity and good reproducibility, demonstrating efficient and stable photoelectric conversion ability of the Ir(III) complex. PMID:25816127

  14. Electrochemical chip-based genomagnetic assay for detection of high-risk human papillomavirus DNA.

    PubMed

    Bartosik, Martin; Durikova, Helena; Vojtesek, Borivoj; Anton, Milan; Jandakova, Eva; Hrstka, Roman

    2016-09-15

    Cervical cancer, being the fourth leading cause of cancer death in women worldwide, predominantly originates from a persistent infection with a high-risk human papillomavirus (HPV). Detection of DNA sequences from these high-risk strains, mostly HPV-16 and HPV-18, represents promising strategy for early screening, which would help to identify women with higher risk of cervical cancer. In developing countries, inadequate screening options lead to disproportionately high mortality rates, making a fast and inexpensive detection schemes highly important. Electrochemical sensors and assays offer an alternative to current methods of detection. We developed an electrochemical-chip based assay, in which target HPV DNA is captured via magnetic bead-modified DNA probes, followed by an antidigoxigenin-peroxidase detection system at screen-printed carbon electrode chips, enabling parallel measurements of eight samples simultaneously. We show sensitive detection in attomoles of HPV DNA, selective discrimination between HPV-16 and HPV-18 and good reproducibility. Most importantly, we show application of the assay into both cancer cell lines and cervical smears from patients. The electrochemical results correlated well with standard methods, making this assay potentially applicable in clinical practice. PMID:27132004

  15. A label-free electrochemical aptasensor based on graphene oxide/double-stranded DNA nanocomposite.

    PubMed

    Li, Yu; Wang, Qi; Zhang, Yuting; Deng, Dongmei; He, Haibo; Luo, Liqiang; Wang, Zhenxin

    2016-09-01

    A novel label-free electrochemical impedance aptasensor based on a gold nanoparticles/double-stranded DNA-graphene (AuNPs/dsDNA-GO) nanocomposite modified glassy carbon electrode was presented for quantitative determination of thrombin. GO was covalently functionalized with dsDNA via a facile amidation process, and then AuNPs were electrodeposited onto the surface of dsDNA-GO. The morphology, conductivity and interaction of the as-prepared nanocomposites were characterized by scanning electron microscopy, cyclic voltammetry, electrochemical impedance spectroscopy (EIS), Raman and Fourier transform infrared spectroscopy. The thrombin-binding aptamer (TBA) was conjugated to AuNPs via gold-thiol chemistry to construct electrochemical aptasensing platform, and the specific recognition between TBA and thrombin was monitored by EIS. Under optimum conditions, thrombin could be quantified in a wide range of 0.1-100nM (R(2)=0.9960) with low detection limit of 0.06nM (S/N=3). PMID:27182650

  16. The non-specific inhibition of enzymes by environmental pollutants: a study of a model system towards the development of electrochemical biosensor arrays.

    PubMed

    Young, S J; Hart, J P; Dowman, A A; Cowell, D C

    2001-12-01

    Previous research has shown that lactate dehydrogenase (LDH) was competitively inhibited by pentachlorophenol (PCP) and a modified assay produced a detection limit of 1 microM (270 microg l(-1)). This work used spectrophotometric rate-determination but in order to move towards biosensor development the selected detection method was electrochemical. The linkage of LDH to lactate oxidase (LOD) provided the electroactive species, hydrogen peroxide. This could be monitored using a screen-printed carbon electrode (SPCE) incorporating the mediator, cobalt phthalocyanine, at a potential of +300 mV (vs. Ag/AgCl). A linked LDH/LOD system was optimised with respect to inhibition by PCP. It was found that the SPCE support material, PVC, acted to reduce inhibition, possibly by combining with PCP. A cellulose acetate membrane removed this effect. Inhibition of the system was greatest at enzyme activities of 5 U ml(-1) LDH and 0.8 U ml(-1) LOD in reactions containing 246 microM pyruvate and 7.5 microM NADPH. PCP detection limits were an EC(10) of 800 nM (213 microg l(-1)) and a minimum inhibition detectable (MID) limit of 650 nM (173 microg l(-1)). The inclusion of a third enzyme, glucose dehydrogenase (GDH), provided cofactor recycling to enable low concentrations of NADPH to be incorporated within the assay. NADPH was reduced from 7.5 to 2 microM. PCP detection limits were obtained for an assay containing 5 U ml(-1) LDH, 0.8 U ml(-1) LOD and 0.1 U ml(-1) GDH with 246 microM pyruvate, 400 mM glucose and 2 microM NADPH. The EC(10) limit was 150 nM (39.9 microg l(-1)) and the MID was 100 nM (26.6 microg l(-1)). The design of the inhibition assays discussed has significance as a model for other enzymes and moves forward the possibility of an electrochemical biosensor array for pollution monitoring. PMID:11679267

  17. Gold coated ZnO nanorod biosensor for glucose detection

    NASA Astrophysics Data System (ADS)

    Bhattacharya, Anuradha; Jain, Chhavi; Rao, V. Padmanapan; Banerjee, S.

    2012-06-01

    Gold coated ZnO nanorod based biosensor has been fabricated for its glucose detecting abilities and compared with that of ZnO nanorod based biosensor. SEM images of electrochemically grown ZnO nanorods show hexagonally grown ZnO nanorods on an ITO substrate. Electrochemical analysis show that gold coated ZnO based biosensors have higher sensitivity, lower limit of detection and a wider linear range for glucose detection. The results demonstrate that gold coated ZnO nanorod based biosensors are a promising material for biosensor applications over single component ZnO nanorod based biosensor.

  18. Electrochemical control of a DNA Holliday Junction nanoswitch by Mg2+ ions.

    PubMed

    Ferapontova, E E; Mountford, C P; Crain, J; Buck, A H; Dickinson, P; Beattie, J S; Ghazal, P; Terry, J G; Walton, A J; Mount, A R

    2008-11-15

    The molecular conformation of a synthetic branched, 4-way DNA Holliday junction (HJ) was electrochemically switched between the open and closed (stacked) conformers. Switching was achieved by electrochemically induced quantitative release of Mg(2+) ions from the oxidised poly(N-methylpyrrole) film (PPy), which contained polyacrylate as an immobile counter anion and Mg(2+) ions as charge compensating mobile cations. This increase in the Mg(2+) concentration screened the electrostatic repulsion between the widely separated arms in the open HJ configuration, inducing switching to the closed conformation. Upon electrochemical reduction of PPy, entrapment of Mg(2+) ions back into the PPy film induced the reverse HJ switching from the closed to open state. The conformational transition was monitored using fluorescence resonance energy transfer (FRET) between donor and acceptor dyes each located at the terminus of one of the arms. The demonstrated electrochemical control of the conformation of the used probe-target HJ complex, previously reported as a highly sequence specific nanodevice for detecting of unlabelled target [Buck, A.H., Campbell, C.J., Dickinson, P., Mountford, C.P., Stoquert, H.C., Terry, J.G., Evans, S.A.G., Keane, L., Su, T.J., Mount, A.R., Walton, A.J., Beattie, J.S., Crain, J., Ghazal, P., 2007. Anal. Chem., 79, 4724-4728], allows the development of electronically addressable DNA nanodevices and label-free gene detection assays. PMID:18541422

  19. Control of electrochemical signals from quantum dots conjugated to organic materials by using DNA structure in an analog logic gate.

    PubMed

    Chen, Qi; Yoo, Si-Youl; Chung, Yong-Ho; Lee, Ji-Young; Min, Junhong; Choi, Jeong-Woo

    2016-10-01

    Various bio-logic gates have been studied intensively to overcome the rigidity of single-function silicon-based logic devices arising from combinations of various gates. Here, a simple control tool using electrochemical signals from quantum dots (QDs) was constructed using DNA and organic materials for multiple logic functions. The electrochemical redox current generated from QDs was controlled by the DNA structure. DNA structure, in turn, was dependent on the components (organic materials) and the input signal (pH). Independent electrochemical signals from two different logic units containing QDs were merged into a single analog-type logic gate, which was controlled by two inputs. We applied this electrochemical biodevice to a simple logic system and achieved various logic functions from the controlled pH input sets. This could be further improved by choosing QDs, ionic conditions, or DNA sequences. This research provides a feasible method for fabricating an artificial intelligence system. PMID:27116705

  20. DNA Hybridization Sensors Based on Electrochemical Impedance Spectroscopy as a Detection Tool

    PubMed Central

    Park, Jin-Young; Park, Su-Moon

    2009-01-01

    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. PMID:22303136

  1. The amplification effect of functionalized gold nanoparticles on the binding of anticancer drug dacarbazine to DNA and DNA bases

    NASA Astrophysics Data System (ADS)

    Shen, Qin; Wang, Xuemei; Fu, Degang

    2008-11-01

    The promising application of functionalized gold nanoparticles to amplify the performance of biosensors and relevant biomolecular recognition processes has been explored in this paper. Our observations illustrate the apparent enhancement effect of the gold nanoparticles on the electrochemical response of the anticancer drug dacarbazine (DTIC) binding to DNA and DNA bases, indicating that these functionalized gold nanoparticles could readily facilitate the specific interactions between DTIC and DNA/DNA bases. This raises the potential valuable applications of these biocompatible nanoparticles in the promising biosensors and biomedical engineering.

  2. Development of an optical biosensor based on surface-enhanced Raman scattering for DNA analysis

    NASA Astrophysics Data System (ADS)

    Yigit, Tugce; Akdogan, Ebru; Karagoz, Isık. Didem; Kahraman, Mehmet

    2016-03-01

    Rapid, accurate and sensitive DNA analysis is critically important for the diagnostic of genetic diseases. The most common method preferred in practice is fluorescence based microarrays to analyze the DNA. However, there exist some disadvantages related to the above-mentioned method such as the overlapping of the fluorescence emission wavelengths that can diminish in the performance of multiplexing, needed to obtain fluorescence spectra from each dye and photo degradation. In this study, a novel SERS based DNA analysis approach, which is Raman active dye-free and independent of SERS substrate properties, is developed. First, the single strand DNA probe is attached to the SERS substrate and half of the complimentary DNA is attached to gold nanoparticles, as well. We hypothesize that in the presence of target DNA, the complimentary DNA coupled colloids will bind to the SERS substrate surface via hybridization of single strand target DNA. To test this hypothesis, we used UV/Vis spectroscopy, atomic for microscopy (AFM) and dynamic light scattering (DLS). DNA analysis is demonstrated by a peak shift of the certain peak of the small molecules attached to the SERS substrate surface instead of SERS spectrum obtained in the presence of target DNA from the Raman reporter molecules. The degree of peak shifting will be used for the quantification of the target DNA in the sample. Plasmonic properties of SERS substrates and reproducibility issues will not be considerable due to the use of peak shifting instead of peak intensity for the qualitative analysis.

  3. An electrochemical biosensor based on nanoporous stainless steel modified by gold and palladium nanoparticles for simultaneous determination of levodopa and uric acid.

    PubMed

    Rezaei, Behzad; Shams-Ghahfarokhi, Leila; Havakeshian, Elaheh; Ensafi, Ali A

    2016-09-01

    In this paper, an electrochemical biosensor based on gold and palladium nano particles-modified nanoporous stainless steel (Au-Pd/NPSS) electrode has been introduced for the simultaneous determination of levodopa (LD) and uric acid (UA). To prepare the electrode, the stainless steel was anodized to fabricate NPSS and then Cu was electrodeposited onto the nanoporous steel by applying the multiple step potential. Finally, the electrode was immersed into a gold and palladium precursor's solution by the atomic ratio of 9:1 to form Au-Pd/NPSS through the galvanic replacement reaction. Morphological aspects, structural properties and the electroanalytical behavior of the Au-Pd/NPSS electrode were studied using field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), electrochemical impedance spectroscopy (EIS) and voltammetric techniques. Also, differential pulse voltammetry (DPV) was used for the simultaneous determination of LD and UA. According to results, the surface of Au-Pd/NPSS electrode contained Au and Pd nanoparticles with an average diameter of 75nm. The electrode acted better than Au/NPSS and Pd/NPSS electrodes for the simultaneous determination of LD and UA, with the peak separation potential of about 220mV. Also, the calibration plot for LD was in two linear concentration ranges of 5.0-10.0 and 10.0-55.0μmolL(-1) and for UA, it was in the range of 100-1200μmolL(-1). The detection limit for LD and UA was 0.2 and 15μmolL(-1), respectively. The modified electrode had a good performance for LD and UA detection in urine, blood serum and levodopa C-Forte tablet. PMID:27343576

  4. Molecular conductance of double-stranded DNA evaluated by electrochemical capacitance spectroscopy.

    PubMed

    Ribeiro, W C; Gonçalves, L M; Liébana, S; Pividori, M I; Bueno, P R

    2016-04-21

    Conductance was measured in two different double stranded DNA (both with 20 bases), the more conducting poly(dG)-poly(dC) (ds-DNAc) and the less conducting poly(dA)-poly(dT) (ds-DNAi), by means of Electrochemical Capacitance Spectroscopy (ECS). The use of the ECS approach, exemplified herein with DNA nanowires, is equally a suitable and time-dependent advantageous alternative for conductance measurement of molecular systems, additionally allowing better understanding of the alignment existing between molecular scale conductance and electron transfer rate. PMID:27074378

  5. Microfluidics and nanoparticles based amperometric biosensor for the detection of cyanobacteria (Planktothrix agardhii NIVA-CYA 116) DNA.

    PubMed

    Ölcer, Zehra; Esen, Elif; Ersoy, Aylin; Budak, Sinan; Sever Kaya, Dilek; Yağmur Gök, Mehmet; Barut, Serkan; Üstek, Duran; Uludag, Yildiz

    2015-08-15

    Some of the cyanobacteria produce protease inhibitor oligopeptides such as cyanopeptolins and cause drinking water contamination; hence, their detection has great importance to monitor the well-being of water sources that is used for human consumption. In the current study, a fast and sensitive nucleic acid biosensor assay has been described where cyanopeptolin coding region of one of the cyanobacteria (Planktothrix agardhii NIVA-CYA 116) genome has been used as target for monitoring of the fresh water resources. A biochip that has two sets of Au electrode arrays, each consist of shared reference/counter electrodes and 3 working electrodes has been used for the assay. The biochip has been integrated to a microfluidics system and all steps of the assay have been performed during the reagent flow to achieve fast and sensitive DNA detection. On-line hybridization of the target on to the capture probe immobilized surface resulted in a very short assay duration with respect to the conventional static assays. The binding of the avidin and enzyme modified Au nanoparticles to the biotinylated detection probe and the subsequent injection of the substrate enabled a real-time amperometric measurement with a detection limit of 6×10(-12) M target DNA (calibration curve r(2)=0.98). The developed assay enables fast and sensitive detection of cyanopeptolin producing cyanobacteria from freshwater samples and hence shows a promising technology for toxic microorganism detection from environmental samples. PMID:25845335

  6. Gold nanoparticle-based lateral flow biosensor for rapid visual detection of Leishmania-specific DNA amplification products.

    PubMed

    Toubanaki, Dimitra K; Athanasiou, Evita; Karagouni, Evdokia

    2016-08-01

    Leishmaniasis is a disease, caused by Leishmania parasites, which infect humans and animals, posing a major social and economic burden worldwide. The need for accurate and sensitive disease diagnosis led to the widespread adoption of PCR amplification. Detection of the amplification products (i.e. gel electrophoresis) require time-consuming protocols performed by trained personnel, with high cost. Aim of the present study was the simplification of PCR product detection, using a nucleic acid lateral flow, combined with functionalized gold nanoparticles. Amplification reactions targeting kinetoplastid DNA of Leishmania spp were performed on canine blood samples and a positive signal was formed as a red test zone. The visual detection was completed in 20min. Extensive optimization enabled the detection of 100fmol of target DNA. Clinical samples of infected dog blood were analyzed with high specificity. Overall, the proposed lateral flow biosensor can be considered an appealing alternative platform for Leishmania-specific amplification products detection with low cost and attractive simplicity. PMID:27255490

  7. An amperometric urea biosensor based on covalent immobilization of urease onto an electrochemically prepared copolymer poly (N-3-aminopropyl pyrrole-co-pyrrole) film.

    PubMed

    Rajesh; Bisht, Vandana; Takashima, Wataru; Kaneto, Keiichi

    2005-06-01

    An amperometric biosensor has been developed for the quantitative determination of urea in aqueous solution. The principle is based on the use of pH-sensitive redox active dissolved hematein molecule. The enzyme, urease (Urs), was covalently immobilized on a conducting copolymer poly (N-3-aminopropyl pyrrole-co-pyrrole) film, electrochemically prepared onto an indium-tin-oxide (ITO)-coated glass plate. The covalent linkage of enzyme and porous morphology of the polymer film lead to high enzyme loading and an increased lifetime stability of the enzyme electrode. Amperometric response was measured as a function of concentration of urea, at fixed bias voltage of 0.0 V vs. Ag/AgCl in a phosphate buffer (pH 7.0). The electrode gives a linear response range of 0.16-5.02 mM for urea in aqueous medium. The response time is 40 s reaching to a 95% steady-state current value, and 80% of the enzyme activity is retained for about 2 months. PMID:15744952

  8. Direct electrochemistry of glucose oxidase at electrochemically reduced graphene oxide-multiwalled carbon nanotubes hybrid material modified electrode for glucose biosensor.

    PubMed

    Mani, Veerappan; Devadas, Balamurugan; Chen, Shen-Ming

    2013-03-15

    Direct electrochemistry of glucose oxidase (GOx) at an electrochemically reduced graphene oxide-multiwalled carbon nanotubes hybrid (ERGO-MWCNT) modified glassy carbon electrode (GCE) has been reported. The π-π stacking interaction operating between the MWCNT and graphene oxide (GO) has been revealed by UV-Vis absorption spectroscopy. GOx was well immobilized onto the ERGO-MWCNT hybrid film, as a result direct electrochemistry of GOx has been achieved. Compared with pristine MWCNT, 2.1 fold higher peak current and very low peak to peak separation (ΔE(p)) of 26 mV were observed at the hybrid film, demonstrating faster electron transfer between GOx and the modified electrode surface. Moreover, the modified film exhibited high electrocatalytic activity towards glucose via reductive detection of oxygen consumption and in the presence of mediator. The proposed biosensor exhibits low detection limit of 4.7 μM with wide linear range of 0.01-6.5mM and acquires excellent storage and operational stabilities. The accurate glucose determination in human blood serum and good recoveries achieved in spiked urine samples revealed their great potential in the practical applications. PMID:22964382

  9. An electrochemical aptamer biosensor based on "gate-controlled" effect using β-cyclodextrin for ultra-sensitive detection of trace mercury.

    PubMed

    Li, Jianping; Sun, Mi; Wei, Xiaoping; Zhang, Lieping; Zhang, Yun

    2015-12-15

    A new strategy for construction an electrochemical biosensor for Hg(2+) analysis with extremely high sensitivity was proposed based on "T-Hg(2+)-T" coordination by aptamers and "gate-controlled" amplification by switch of the channels of the probe. SH-β-cyclodextrin was self-assembled on the gold electrode to form an orderly arranged molecularly layer with interspaces among β-CDs; thionine labeled aptamer was then linked on the assembled β-CD. When Hg(2+) was added, the aptamer combined with Hg(2+) and formed "T-Hg(2+)-T" structure, which caused the aptamer folded and the labeled thionine covered the interspaces to block off the channel for probe entrance. The oxidative current of probe decreased, which provide the basis for the determination of Hg(2+). With the gate-controlled amplification, the changes of trace amounts of Hg(2+) will produce great changes of the probe current. The sensor exhibited significantly higher sensitivity with a detection limit of 5.0×10(-15) mol/L, which is lower than other reported methods. PMID:26164015

  10. Targeting CpG DNA to screen and isolate anti-sepsis fraction and monomers from traditional Chinese herbs using affinity biosensor technology.

    PubMed

    Liu, Xin; Cheng, Juan; Zheng, Xinchuang; Chen, Yiguo; Wu, Chong; Li, Bin; Fu, Jianfeng; Cao, Hongwei; Lu, Yongling; Li, Jun; Zheng, Jiang; Zhou, Hong

    2009-08-01

    Bacterial DNA/CpG DNA is recognized as a key molecule during the pathogenesis of sepsis. Therefore, preventing CpG DNA from binding to its receptor is considered as the most promising strategy. In the present experiments, Radix et Rhizoma Rhei had the highest CpG DNA-binding ability among the seventy-eight traditional Chinese herbs. After the isolation of silica gel chromatography and high performance liquid chromatography (HPLC) and evaluation with affinity biosensor, the active fraction was confirmed and named Fraction D. It was found that in vitro, Fraction D bound to both CpG DNA and lipid A with high affinity, and strongly inhibited LPS- and CpG DNA-induced TNF-alpha release from RAW264.7 cells in a dose-dependent manner. Furthermore, Fraction D reduced the expression of TLR9 mRNA up-regulated by CpG DNA. In vivo, Fraction D protected mice challenged with lethal heat-killed E. coli. Using HPLC method, two monomers with high affinity for CpG DNA were isolated and identified as rhein and emodin. Rhein could significantly reduce CpG DNA- and LPS-induced TNF-alpha release, but emodin only reduced CpG DNA-induced TNF-alpha release. Rhein in combination with emodin could play synergistic inhibitory effect on both CpG DNA and LPS-induced TNF-alpha release, which contributed to the bioactivity of Fraction D. In conclusion, we successfully established the platform to screen anti-CpG DNA components of traditional Chinese herbs using affinity biosensor technology, got active Fraction D from Radix et Rhizoma Rhei and determined rhein and emodin as the main bioactive ingredients in Fraction D. PMID:19376273

  11. New metal based drugs: Spectral, electrochemical, DNA-binding, surface morphology and anticancer activity properties

    NASA Astrophysics Data System (ADS)

    Çeşme, Mustafa; Gölcü, Aysegul; Demirtaş, Ibrahim

    2015-01-01

    The NSAID piroxicam (PRX) drug was used for complex formation reactions with Cu(II), Zn(II) and Pt(II) metal salts have been synthesized. Then, these complexes have been characterized by spectroscopic and analytical techniques. Thermal behavior of the complexes were also investigated. The electrochemical properties of all complexes have been investigated by cyclic voltammetry (CV) using glassy carbon electrode. The biological activity of the complexes has been evaluated by examining their ability to bind to fish sperm double strand DNA (FSFSdsDNA) with UV spectroscopy. UV studies of the interaction of the PRX and its complexes with FSdsDNA have shown that these compounds can bind to FSdsDNA. The binding constants of the compounds with FSdsDNA have also been calculated. The morphology of the FSdsDNA, PRX, metal ions and metal complexes has been investigated by scanning electron microscopy (SEM). To get the SEM images, the interaction of compounds with FSdsDNA has been studied by means of differential pulse voltammetry (DPV) at FSdsDNA modified pencil graphite electrode (PGE). The decrease in intensity of the guanine oxidation signals has been used as an indicator for the interaction mechanism. The effect of proliferation PRX and complexes were examined on the HeLA and C6 cells using real-time cell analyzer with four different concentrations.

  12. Label-free, disposable fiber-optic biosensors for DNA hybridization detection.

    PubMed

    Yin, Ming-jie; Wu, Chuang; Shao, Li-yang; Chan, Wing Kin Edward; Zhang, A Ping; Lu, Chao; Tam, Hwa-yaw

    2013-04-01

    A novel and highly sensitive fiber-optic DNA sensor based on a thin-core fiber modal interferometer (TCFMI) is demonstrated by using a layer-by-layer (LbL) self-assembly technology. Poly(ethylenimine) (PEI), poly(acrylic acid) (PAA) and single-stranded DNA (ssDNA) were used for the preparation of a polyelectrolyte multilayer film for DNA detection. The film thickness was measured through a surface profilometer. The surface morphologies of (PEI/PAA)4, (PEI/PAA)4.5 and (PEI/PAA)4(PEI/DNA)1 multilayer films were characterized by atomic force microscopy. The fabricated DNA sensors were tested with different types of target ssDNA solutions with a concentration of 1 μM. The results show that the sensitivity of the TCFMI-based ssDNA sensor is 0.27 nm/matched-base at the concentration of 1 μM and can even distinguish the number of matched bases of ssDNA chains. PMID:23397583

  13. Development of a lab-on-chip electrochemical biosensor for water quality analysis based on microalgal photosynthesis.

    PubMed

    Tsopela, A; Laborde, A; Salvagnac, L; Ventalon, V; Bedel-Pereira, E; Séguy, I; Temple-Boyer, P; Juneau, P; Izquierdo, R; Launay, J

    2016-05-15

    The present work was dedicated to the development of a lab-on-chip device for water toxicity analysis and more particularly herbicide detection in water. It consists in a portable system for on-site detection composed of three-electrode electrochemical microcells, integrated on a fluidic platform constructed on a glass substrate. The final goal is to yield a system that gives the possibility of conducting double, complementary detection: electrochemical and optical and therefore all materials used for the fabrication of the lab-on-chip platform were selected in order to obtain a device compatible with optical technology. The basic detection principle consisted in electrochemically monitoring disturbances in metabolic photosynthetic activities of algae induced by the presence of Diuron herbicide. Algal response, evaluated through oxygen (O2) monitoring through photosynthesis was different for each herbicide concentration in the examined sample. A concentration-dependent inhibition effect of the herbicide on photosynthesis was demonstrated. Herbicide detection was achieved through a range (blank - 1 µM Diuron herbicide solution) covering the limit of maximum acceptable concentration imposed by Canadian government (0.64 µM), using a halogen white light source for the stimulation of algal photosynthetic apparatus. Superior sensitivity results (limit of detection of around 0.1 µM) were obtained with an organic light emitting diode (OLED), having an emission spectrum adapted to algal absorption spectrum and assembled on the final system. PMID:26749098

  14. Preparation and characterization of three dimensional graphene foam supported platinum-ruthenium bimetallic nanocatalysts for hydrogen peroxide based electrochemical biosensors.

    PubMed

    Kung, Chih-Chien; Lin, Po-Yuan; Buse, Frederick John; Xue, Yuhua; Yu, Xiong; Dai, Liming; Liu, Chung-Chiun

    2014-02-15

    The large surface, the excellent dispersion and the high degrees of sensitivity of bimetallic nanocatalysts were the attractive features of this investigation. Graphene foam (GF) was a three dimensional (3D) porous architecture consisting of extremely large surface and high conductive pathways. In this study, 3D GF was used incorporating platinum-ruthenium (PtRu) bimetallic nanoparticles as an electrochemical nanocatalyst for the detection of hydrogen peroxide (H2O2). PtRu/3D GF nanocatalyst exhibited a remarkable performance toward electrochemical oxidation of H2O2 without any additional mediator showing a high sensitivity (1023.1 µA mM(-1)cm(-2)) and a low detection limit (0.04 µM) for H2O2. Amperometric results demonstrated that GF provided a promising platform for the development of electrochemical sensors in biosensing and PtRu/3D GF nanocatalyst possessed the excellent catalytic activity toward the H2O2 detection. A small particle size and a high degree of the dispersion in obtaining of large active surface area were important for the nanocatalyst for the best H2O2 detection in biosensing. Moreover, potential interference by ascorbic acid and uric acid appeared to be negligible. PMID:24012804

  15. Electrochemical detection of DNA 3'-phosphatases based on surface-extended DNA nanotail strategy.

    PubMed

    Wu, Dan; Li, Chao; Hu, Xiaolu; Mao, Xiaoxia; Li, Genxi

    2016-06-14

    Determination of DNA dephosphorylation is of great value due to its vital role in many cellular processes. Here we report a surface-extended DNA nanotail strategy for simple and ultrasensitive detection of DNA 3'-phosphatases by terminal deoxynucleotidyl transferase (TdT) mediated signal amplification. In this work, DNA probes labeled with thiols at their 5' terminals and phosphoryls at 3' terminals are immobilized on gold electrode and are used as substrates for DNA 3'-phosphatases, taking T4 polynucleotide kinase phosphatase (T4PNKP) as an example. T4PNKP can catalyze the dephosphorylation reaction of the substrate DNA, followed by the formation of a long DNA strand by TdT on its 3' terminal hydroxyl, leading to an evident chronocoulometry signal enhancement. The proposal presents a considerable analytical performance with low detection limit and wide linear range, making it promise to be applied in the fields of DNA dephosphorylation related processes, drug discovery, and clinical diagnostics. PMID:27181641

  16. Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression

    PubMed Central

    Geng, Deyu; Zhang, Zhixia; Guo, Huarong

    2012-01-01

    p21CIP1/WAF1 is a p53-target gene in response to cellular DNA damage. Here we report the development of a fish cell biosensor system for high throughput genotoxicity detection of new drugs, by stably integrating two reporter plasmids of pGL3-p21-luc (human p21 promoter linked to firefly luciferase) and pRL-CMV-luc (CMV promoter linked to Renilla luciferase) into marine flatfish flounder gill (FG) cells, referred to as p21FGLuc. Initial validation of this genotoxicity biosensor system showed that p21FGLuc cells had a wild-type p53 signaling pathway and responded positively to the challenge of both directly acting genotoxic agents (bleomycin and mitomycin C) and indirectly acting genotoxic agents (cyclophosphamide with metabolic activation), but negatively to cyclophosphamide without metabolic activation and the non-genotoxic agents ethanol and D-mannitol, thus confirming a high specificity and sensitivity, fast and stable response to genotoxic agents for this easily maintained fish cell biosensor system. This system was especially useful in the genotoxicity detection of Di(2-ethylhexyl) phthalate (DEHP), a rodent carcinogen, but negatively reported in most non-mammalian in vitro mutation assays, by providing a strong indication of genotoxicity for DEHP. A limitation for this biosensor system was that it might give false positive results in response to sodium butyrate and any other agents, which can trans-activate the p21 gene in a p53-independent manner. PMID:25585933

  17. Label free detection of DNA hybridization by refractive index tapered fiber biosensor

    NASA Astrophysics Data System (ADS)

    Zibaii, M. I.; Latifi, H.; Ghanati, E.; Gholami, M.; Hosseini, S. M.

    2010-04-01

    We demonstrate a simple refractive index sensor (RI) sensing system based on a biconical tapered optical fiber (BTOF), which is fabricated by heat pulling method, utilizing a CO2 laser. In this work we explore the application of these sensors for the detection of label free single stranded DNA (ssDNA) in real time. During the experiment, the target ssDNA did not need to be labeled with a fluorescent tag, which is expensive and complicated. The change in output optical transmission of the tapered fiber was recorded for Poly-L-Lysine (PLL) coating, ssDNA probe immobilization and hybridization. The result indicated that due to the hybridization with the complementary target ssDNA on the tapered surface, the RI of surrounding medium changes which leads to changes in the characteristics of the tapered region and change in the output power of the sensor.

  18. Molecular conductance of double-stranded DNA evaluated by electrochemical capacitance spectroscopy

    NASA Astrophysics Data System (ADS)

    Ribeiro, W. C.; Gonçalves, L. M.; Liébana, S.; Pividori, M. I.; Bueno, P. R.

    2016-04-01

    Conductance was measured in two different double stranded DNA (both with 20 bases), the more conducting poly(dG)-poly(dC) (ds-DNAc) and the less conducting poly(dA)-poly(dT) (ds-DNAi), by means of Electrochemical Capacitance Spectroscopy (ECS). The use of the ECS approach, exemplified herein with DNA nanowires, is equally a suitable and time-dependent advantageous alternative for conductance measurement of molecular systems, additionally allowing better understanding of the alignment existing between molecular scale conductance and electron transfer rate.Conductance was measured in two different double stranded DNA (both with 20 bases), the more conducting poly(dG)-poly(dC) (ds-DNAc) and the less conducting poly(dA)-poly(dT) (ds-DNAi), by means of Electrochemical Capacitance Spectroscopy (ECS). The use of the ECS approach, exemplified herein with DNA nanowires, is equally a suitable and time-dependent advantageous alternative for conductance measurement of molecular systems, additionally allowing better understanding of the alignment existing between molecular scale conductance and electron transfer rate. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr01076h

  19. Electrochemical detection of Francisella tularensis genomic DNA using solid-phase recombinase polymerase amplification.

    PubMed

    del Río, Jonathan Sabaté; Yehia Adly, Nouran; Acero-Sánchez, Josep Lluis; Henry, Olivier Y F; O'Sullivan, Ciara K

    2014-04-15

    Solid-phase isothermal DNA amplification was performed exploiting the homology protein recombinase A (recA). The system was primarily tested on maleimide activated microtitre plates as a proof-of-concept and later translated to an electrochemical platform. In both cases, forward primer for Francisella tularensis holarctica genomic DNA was surface immobilised via a thiol or an amino moiety and then elongated during the recA mediated amplification, carried out in the presence of specific target sequence and reverse primers. The formation of the subsequent surface tethered amplicons was either colorimetrically or electrochemically monitored using a horseradish peroxidase (HRP)-labelled DNA secondary probe complementary to the elongated strand. The amplification time was optimised to amplify even low amounts of DNA copies in less than an hour at a constant temperature of 37°C, achieving a limit of detection of 1.3×10(-13) M (4×10(6) copies in 50 μL) for the colorimetric assay and 3.3×10(-14) M (2×10(5) copies in 10 μL) for the chronoamperometric assay. The system was demonstrated to be highly specific with negligible cross-reactivity with non-complementary targets or primers. PMID:24334283

  20. Novel trends in affinity biosensors: current challenges and perspectives

    NASA Astrophysics Data System (ADS)

    Arugula, Mary A.; Simonian, Aleksandr

    2014-03-01

    Molecular biorecognition processes facilitate physical and biochemical interactions between molecules in all crucial metabolic pathways. Perhaps the target analyte and the biorecognition element interactions have the most impactful use in biosensing applications. Traditional analytical sensing systems offer excellent biorecognition elements with the ability to detect and determine the presence of analytes. High affinity antibodies and DNA play an important role in the development of affinity biosensors based on electrochemical, optical and mass sensitive approaches. Advancements in this area routinely employ labels, label free, nanoparticles, multifunctional matrices, carbon nanotubes and other methods to meet the requirements of its own application. However, despite increasing affinity ceilings for conventional biosensors, the field draws back in meeting specifically important demands, such as long-term stability, ultrasensitivity, rapid detection, extreme selectivity, strong biological base, calibration, in vivo measurements, regeneration, satisfactory performance and ease of production. Nevertheless, recent efforts through this line have produced novel high-tech nanosensing systems such as ‘aptamers’ and ‘phages’ which exhibit high-throughput sensing. Aptamers and phages are powerful tools that excel over antibodies in sensibility, stability, multi-detection, in vivo measurements and regeneration. Phages are superior in stability, screening for affinity-based target molecules ranging from small to proteins and even cells, and easy production. In this review, we focus mainly on recent developments in affinity-based biosensors such as immunosensors, DNA sensors, emphasizing aptasensors and phage-based biosensors basing on novel electrochemical, optical and mass sensitive detection techniques. We also address enzyme inhibition-based biosensors and the current problems associated with the above sensors and their future perspectives.

  1. A novel signal-on electrochemical DNA sensor based on target catalyzed hairpin assembly strategy.

    PubMed

    Qian, Yong; Tang, Daoquan; Du, Lili; Zhang, Yanzhuo; Zhang, Lixian; Gao, Fenglei

    2015-02-15

    We describe a novel signal-on electrochemical DNA (E-DNA) sensing platform based on target-catalyzed hairpin assembly. The thiolated modified molecular beacon 1 (MB1) was first immobilized onto the Au electrode (GE) surface and then target DNA hybridized to the MB1, the opened MB1 assembled with the ferrocene (Fc)-labeled molecular beacon 2 to displace the target DNA, which became available for the next cycle of MB1-target hybridization. Moreover, Fc was confined close to the GE surface for efficient electron transfer, resulting in a current signal. Eventually, each target strand went through many cycles, resulting in numerous Fcs confining close to the GE, which leaded to the current of Fc dramatically increase. The observed signal gain was sufficient to achieve a demonstrated detection limit of 0.74 fM, with a wide linear dynamic range from 10(-15) to 10(-10)M and discriminated mismatched DNA from perfect matched target DNA with a high selectivity. Thus, the proposed E-DNA sensor would have a wide range of sensor applications because it is enzyme-free and simple to perform. PMID:25218101

  2. DNA biosensors implemented on PNA-functionalized microstructured optical fibers Bragg gratings

    NASA Astrophysics Data System (ADS)

    Candiani, A.; Giannetti, S.; Cucinotta, A.; Bertucci, A.; Manicardi, A.; Konstantaki, M.; Margulis, W.; Pissadakis, S.; Corradini, R.; Selleri, S.

    2013-05-01

    A novel DNA sensing platform based on a Peptide Nucleic Acid - functionalized Microstructured Optical Fibers gratings has been demonstrated. The inner surface of different MOFs has been functionalized using PNA probes, OligoNucleotides mimic that are well suited for specific DNA target sequences detection. The hybrid sensing systems were tested for optical DNA detection of targets of relevance in biomedical application, using the cystic fibrosis gene mutation, and food-analysis, using the genomic DNA from genetic modified organism soy flour. After the solutions of DNA molecules has been infiltrated inside the fibers capillaries and hybridization has occurred, oligonucleotidefunctionalized gold nanoparticles were infiltrated and used to form a sandwich-like system to achieve signal amplification. Spectral measurements of the reflected signal reveal a clear wavelength shift of the reflected modes when the infiltrated complementary DNA matches with the PNA probes placed on the inner fiber surface. Measurements have also been made using the mismatched DNA solution for the c, containing a single nucleotide polymorphism, showing no significant changes in the reflected spectrum. Several experiments have been carried out demonstrating the reproducibility of the results and the high selectivity of the sensors, showing the simplicity and the potential of this approach.

  3. High-Capacity Conductive Nanocellulose Paper Sheets for Electrochemically Controlled Extraction of DNA Oligomers

    PubMed Central

    Razaq, Aamir; Nyström, Gustav; Strømme, Maria; Mihranyan, Albert; Nyholm, Leif

    2011-01-01

    Highly porous polypyrrole (PPy)-nanocellulose paper sheets have been evaluated as inexpensive and disposable electrochemically controlled three-dimensional solid phase extraction materials. The composites, which had a total anion exchange capacity of about 1.1 mol kg−1, were used for extraction and subsequent release of negatively charged fluorophore tagged DNA oligomers via galvanostatic oxidation and reduction of a 30–50 nm conformal PPy layer on the cellulose substrate. The ion exchange capacity, which was, at least, two orders of magnitude higher than those previously reached in electrochemically controlled extraction, originated from the high surface area (i.e. 80 m2 g−1) of the porous composites and the thin PPy layer which ensured excellent access to the ion exchange material. This enabled the extractions to be carried out faster and with better control of the PPy charge than with previously employed approaches. Experiments in equimolar mixtures of (dT)6, (dT)20, and (dT)40 DNA oligomers showed that all oligomers could be extracted, and that the smallest oligomer was preferentially released with an efficiency of up to 40% during the reduction of the PPy layer. These results indicate that the present material is very promising for the development of inexpensive and efficient electrochemically controlled ion-exchange membranes for batch-wise extraction of biomolecules. PMID:22195031

  4. High-sensitivity DNA biosensor based on optical fiber taper interferometer coated with conjugated polymer tentacle.

    PubMed

    Huang, Yunyun; Tian, Zhuang; Sun, Li-Peng; Sun, Dandan; Li, Jie; Ran, Yang; Guan, Bai-Ou

    2015-10-19

    A sensitive bio-probe to in situ detect unlabeled single-stranded DNA targets based on optical microfiber taper interferometer coated by a high ordered pore arrays conjugated polymer has been presented. The polymer coating serves as tentacles to catch single-stranded DNA molecules by π-π conjugated interaction and varies the surface refractive index of the optical microfiber. The microfiber taper interferometer translates the refractive index information into wavelength shift of the interference fringe. The sensor exhibits DNA concentration sensitivity of 2.393 nm/log M and the lowest detection ability of 10(-10) M or even lower. PMID:26480357

  5. Electrochemical Assay for the Signal-on Detection of Human DNA Methyltransferase Activity

    PubMed Central

    Muren, Natalie B.; Barton, Jacqueline K.

    2013-01-01

    Strategies to detect human DNA methyltransferases are needed, given that aberrant methylation by these enzymes is associated with cancer initiation and progression. Here we describe a non-radioactive, antibody-free, electrochemical assay in which methyltransferase activity on DNA-modified electrodes confers protection from restriction for signal-on detection. We implement this assay with a multiplexed chip platform and show robust detection of both bacterial (SssI) and human (Dnmt1) methyltransferase activity. Essential to work with human methyltransferases, our unique assay design allows activity measurements on both unmethylated and hemimethylated DNA substrates. We validate this assay by comparison with a conventional radioactive method. The advantages of electrochemistry over radioactivity and fluorescence make this assay an accessible and promising new approach for the sensitive, label-free detection of human methyltransferase activity. PMID:24164112

  6. Ferrocene-functionalized 4-(2,5-Di(thiophen-2-yl)-1H-pyrrol-1-yl)aniline: a novel design in conducting polymer-based electrochemical biosensors.

    PubMed

    Ayranci, Rukiye; Demirkol, Dilek Odaci; Ak, Metin; Timur, Suna

    2015-01-01

    Herein, we report a novel ferrocenyldithiophosphonate functional conducting polymer and its use as an immobilization matrix in amperometric biosensor applications. Initially, 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)amidoferrocenyldithiophosphonate was synthesized and copolymerized with 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)benzenamine at graphite electrodes. The amino groups on the polymer were utilized for covalent attachment of the enzyme glucose oxidase. Besides, ferrocene on the backbone was used as a redox mediator during the electrochemical measurements. Prior to the analytical characterization, optimization studies were carried out. The changes in current signals at +0.45 V were proportional to glucose concentration from 0.5 to 5.0 mM. Finally, the resulting biosensor was applied for glucose analysis in real samples and the data were compared with the spectrophotometric Trinder method. PMID:25591169

  7. Ferrocene-Functionalized 4-(2,5-Di(thiophen-2-yl)-1H-pyrrol-1-yl)aniline: A Novel Design in Conducting Polymer-Based Electrochemical Biosensors

    PubMed Central

    Ayranci, Rukiye; Demirkol, Dilek Odaci; Ak, Metin; Timur, Suna

    2015-01-01

    Herein, we report a novel ferrocenyldithiophosphonate functional conducting polymer and its use as an immobilization matrix in amperometric biosensor applications. Initially, 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)amidoferrocenyldithiophosphonate was synthesized and copolymerized with 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)benzenamine at graphite electrodes. The amino groups on the polymer were utilized for covalent attachment of the enzyme glucose oxidase. Besides, ferrocene on the backbone was used as a redox mediator during the electrochemical measurements. Prior to the analytical characterization, optimization studies were carried out. The changes in current signals at +0.45 V were proportional to glucose concentration from 0.5 to 5.0 mM. Finally, the resulting biosensor was applied for glucose analysis in real samples and the data were compared with the spectrophotometric Trinder method. PMID:25591169

  8. Study of concentration of HPV DNA probe immobilization for cervical cancer detection based IDE biosensor

    NASA Astrophysics Data System (ADS)

    Roshila, M. L.; Hashim, U.; Azizah, N.

    2016-07-01

    This paper mainly illustrates regarding the detection process of Human Papillomavirus (HPV) DNA probe. HPV is the most common virus that infected to human by a sexually transmitted virus. The most common high-risk HPV are 16 and 18. Interdigitated electrode (IDE) device used as based of Titanium Dioxide (TiO2) acts as inorganic surface, where by using APTES as a linker between inorganic surface and organic surface. A strategy of rapid and sensitive for the HPV detection was proposed by integrating simple DNA extraction with a gene of DNA. The extraction of the gene of DNA will make an efficiency of the detection process. It will depend on the sequence of the capture probes and the way to support their attached. The fabrication, surface modification, immobilization and hybridization processes are characterized by current voltage (I-V) measurement by using KEITHLEY 6487. This strategy will perform a good sensitivity of HPV detection.

  9. Electrochemical characterization of thin film electrodes toward developing a DNA transistor.

    PubMed

    Harrer, Stefan; Ahmed, Shafaat; Afzali-Ardakani, Ali; Luan, Binquan; Waggoner, Philip S; Shao, Xiaoyan; Peng, Hongbo; Goldfarb, Dario L; Martyna, Glenn J; Rossnagel, Stephen M; Deligianni, Lili; Stolovitzky, Gustavo A

    2010-12-21

    The DNA-Transistor is a device designed to control the translocation of single-stranded DNA through a solid-state nanopore. Functionality of the device is enabled by three electrodes exposed to the DNA-containing electrolyte solution within the pore and the application of a dynamic electrostatic potential well between the electrodes to temporarily trap a DNA molecule. Optimizing the surface chemistry and electrochemical behavior of the device is a necessary (but by no means sufficient) step toward the development of a functional device. In particular, effects to be eliminated are (i) electrochemically induced surface alteration through corrosion or reduction of the electrode surface and (ii) formation of hydrogen or oxygen bubbles inside the pore through water decomposition. Even though our motivation is to solve problems encountered in DNA transistor technology, in this paper we report on generic surface chemistry results. We investigated a variety of electrode-electrolyte-solvent systems with respect to their capability of suppressing water decomposition and maintaining surface integrity. We employed cyclic voltammetry and long-term amperometry as electrochemical test schemes, X-ray photoelectron spectroscopy, atomic force microscopy, and scanning, as well as transmission electron microscopy as analytical tools. Characterized electrode materials include thin films of Ru, Pt, nonstoichiometric TiN, and nonstoichiometric TiN carrying a custom-developed titanium oxide layer, as well as custom-oxidized nonstoichiometric TiN coated with a monolayer of hexadecylphosphonic acid (HDPA). We used distilled water as well as aqueous solutions of poly(ethylene glycol) (PEG-300) and glycerol as solvents. One millimolar KCl was employed as electrolyte in all solutions. Our results show that the HDPA-coated custom-developed titanium oxide layer effectively passivates the underlying TiN layer, eliminating any surface alterations through corrosion or reduction within a voltage

  10. DNA-Redox Cation Interaction Improves the Sensitivity of an Electrochemical Immunosensor for Protein Detection

    PubMed Central

    Li, Ping; Ge, Bixia; Ou, Lily M.-L.; Yao, Zhihui; Yu, Hua-Zhong

    2015-01-01

    A simple DNA-redox cation interaction enhancement strategy has been developed to improve the sensitivity of electrochemical immunosensors for protein detection. Instead of labeling with fluorophores or redox-active groups, the detection antibodies were tethered with DNA single strands. Based on the electrostatic interaction between redox cations ([Ru(NH3)6]3+) and negatively charged DNA backbone, enhanced electrochemical signals were obtained. Human chorionic gonadotropin (hCG) detection has been performed as a trial analysis. A linear response range up to the concentration of 25 mIU/mL and a detection limit of 1.25 mIU/mL have been achieved, both are comparable with the ultrasensitive enzyme-linked immunosorbent assay (ELISA) tests. The method also shows great selectivity towards hCG over other hormones such as thyroid stimulating hormone (TSH) and follicle stimulating hormone (FSH). By and large, our approach bears the merits of cost effectiveness and simplicity of instrumentation in comparison with conventional optical detection methods. PMID:26307986

  11. Sensitive and selective real-time electrochemical monitoring of DNA repair (Presentation Recording)

    NASA Astrophysics Data System (ADS)

    Slinker, Jason D.; McWilliams, Marc; Anka, Fadwa; Balkus, Kenneth

    2015-10-01

    Unrepaired DNA damage can lead to mutation, cancer, and death of cells or organisms. However, due to the subtlety of DNA damage, it is difficult to sense the repair of damage products with high selectivity and sensitivity. Here, we show sensitive and selective electrochemical sensing of the repair activity of 8-oxoguanine and uracil glycosylases within DNA monolayers on gold by multiplexed analysis with silicon chips and low-cost electrospun nanofibers. Our approach involves comparing the electrochemical signal of redox probe modified monolayers containing the defect versus the rational control of defect-free monolayers. We find sequence-specific sensitivity thresholds on the order of femtomoles of proteins and dynamic ranges of over two orders of magnitude for each target. For 8-oxoguanine repair, temperature-dependent kinetics are extracted, showing exponential signal loss with time constants of seconds. Electrospun fibers are shown to behave similarly to conventional gold-on-silicon devices, showing the potential of these low-cost devices for sensing applications.

  12. Sample preparation methods for quantitative detection of DNA by molecular assays and marine biosensors.

    PubMed

    Cox, Annie M; Goodwin, Kelly D

    2013-08-15

    The need for quantitative molecular methods is growing in environmental, food, and medical fields but is hindered by low and variable DNA extraction and by co-extraction of PCR inhibitors. DNA extracts from Enterococcus faecium, seawater, and seawater spiked with E. faecium and Vibrio parahaemolyticus were tested by qPCR for target recovery and inhibition. Conventional and novel methods were tested, including Synchronous Coefficient of Drag Alteration (SCODA) and lysis and purification systems used on an automated genetic sensor (the Environmental Sample Processor, ESP). Variable qPCR target recovery and inhibition were measured, significantly affecting target quantification. An aggressive lysis method that utilized chemical, enzymatic, and mechanical disruption enhanced target recovery compared to commercial kit protocols. SCODA purification did not show marked improvement over commercial spin columns. Overall, data suggested a general need to improve sample preparation and to accurately assess and account for DNA recovery and inhibition in qPCR applications. PMID:23790450

  13. A upconversion luminescene biosensor based on dual-signal amplification for the detection of short DNA species of c-erbB-2 oncogene

    PubMed Central

    Lan, Jianming; Liu, Yingxin; Li, Li; Wen, Fadi; Wu, Fang; Han, Zhizhong; Sun, Weiming; Li, Chunyan; Chen, Jinghua

    2016-01-01

    High-sensitivity detection of trace amounts of c-erbB-2 oncogene was reported to be equal to or surpass the ability of CA 15-3 for early diagnosis and/or follow-up recurrent screening of breast cancer. Therefore, in the current study, by using upconversion nanoparticles (UCNPs), rare earth-doped NaYF4:Yb3+/Er3+ as the luminescent labels, a upconversion luminescent (UCL) biosensor based on dual-signal amplification of exonuclease III (ExoIII)-assisted target cycles and long-range self-assembly DNA concatamers was developed for the detection of c-erbB-2 oncogene. The proposed biosensor exhibited ultrasensitive detection with limit as low as 40 aM, which may express the potential of being used in trace analysis of c-erbB-2 oncogene and early diagnosis of breast cancer. PMID:27098295

  14. An electrochemical biosensor for rapid detection of E. coli O157:H7 with highly efficient bi-functional glucose oxidase-polydopamine nanocomposites and Prussian blue modified screen-printed interdigitated electrodes.

    PubMed

    Xu, Meng; Wang, Ronghui; Li, Yanbin

    2016-09-21

    The presence of pathogenic bacteria in foods has always been a great threat to the wellbeing of people and the revenue of food manufacturers. Therefore, the demand for advanced detection methods that can sensitively and rapidly detect these pathogens has been of great importance. This study reports an electrochemical biosensor for rapid detection of E. coli O157:H7 with the integration of bifunctional glucose oxidase (GOx)-polydopamine (PDA) based polymeric nanocomposites (PMNCs) and Prussian blue (PB) modified screen-printed interdigitated microelectrodes (SP-IDMEs). The core-shell magnetic beads (MBs)-GOx@PDA PMNCs were first synthesized by the self-polymerization of dopamine (DA). Gold nanoparticles (AuNPs) were dispersed on the surface of PMNCs through biochemical synthesis to achieve further highly efficient adsorption of antibodies (ABs) and GOx. The final product ABs/GOxext/AuNPs/MBs-GOx@PDA PMNCs served as the carrier to separate target bacteria from food matrices as well as the amplifier for electrochemical measurement. The unbound PMNCs were separated by a filtration step and transferred into glucose solution to allow the enzymatic reaction to occur. The change of the current response was measured with an electrochemical detector using PB-modified SP-IDMEs. The constructed biosensor has been proven to be able to detect E. coli O157:H7 with the detection limit of 10(2) cfu ml(-1). The bifunctional PMNCs contain a high load of enzyme and can optimally utilize the binding sites on bacterial cells, which efficiently amplify the signals for measurement. The biosensor in this study exhibited good specificity, reproducibility, and stability and is expected to have a great impact on applications in the detection of foodborne pathogens. PMID:27358917

  15. Multiplex electrochemical DNA platform for femtomolar-level quantification of genetically modified soybean.

    PubMed

    Manzanares-Palenzuela, C Lorena; de-los-Santos-Álvarez, Noemí; Lobo-Castañón, María Jesús; López-Ruiz, Beatriz

    2015-06-15

    Current EU regulations on the mandatory labeling of genetically modified organisms (GMOs) with a minimum content of 0.9% would benefit from the availability of reliable and rapid methods to detect and quantify DNA sequences specific for GMOs. Different genosensors have been developed to this aim, mainly intended for GMO screening. A remaining challenge, however, is the development of genosensing platforms for GMO quantification, which should be expressed as the number of event-specific DNA sequences per taxon-specific sequences. Here we report a simple and sensitive multiplexed electrochemical approach for the quantification of Roundup-Ready Soybean (RRS). Two DNA sequences, taxon (lectin) and event-specific (RR), are targeted via hybridization onto magnetic beads. Both sequences are simultaneously detected by performing the immobilization, hybridization and labeling steps in a single tube and parallel electrochemical readout. Hybridization is performed in a sandwich format using signaling probes labeled with fluorescein isothiocyanate (FITC) or digoxigenin (Dig), followed by dual enzymatic labeling using Fab fragments of anti-Dig and anti-FITC conjugated to peroxidase or alkaline phosphatase, respectively. Electrochemical measurement of the enzyme activity is finally performed on screen-printed carbon electrodes. The assay gave a linear range of 2-250 pM for both targets, with LOD values of 650 fM (160 amol) and 190 fM (50 amol) for the event-specific and the taxon-specific targets, respectively. Results indicate that the method could be applied for GMO quantification below the European labeling threshold level (0.9%), offering a general approach for the rapid quantification of specific GMO events in foods. PMID:25590971

  16. Optical biosensors

    PubMed Central

    Damborský, Pavel; Švitel, Juraj

    2016-01-01

    Optical biosensors represent the most common type of biosensor. Here we provide a brief classification, a description of underlying principles of operation and their bioanalytical applications. The main focus is placed on the most widely used optical biosensors which are surface plasmon resonance (SPR)-based biosensors including SPR imaging and localized SPR. In addition, other optical biosensor systems are described, such as evanescent wave fluorescence and bioluminescent optical fibre biosensors, as well as interferometric, ellipsometric and reflectometric interference spectroscopy and surface-enhanced Raman scattering biosensors. The optical biosensors discussed here allow the sensitive and selective detection of a wide range of analytes including viruses, toxins, drugs, antibodies, tumour biomarkers and tumour cells. PMID:27365039

  17. Optical biosensors.

    PubMed

    Damborský, Pavel; Švitel, Juraj; Katrlík, Jaroslav

    2016-06-30

    Optical biosensors represent the most common type of biosensor. Here we provide a brief classification, a description of underlying principles of operation and their bioanalytical applications. The main focus is placed on the most widely used optical biosensors which are surface plasmon resonance (SPR)-based biosensors including SPR imaging and localized SPR. In addition, other optical biosensor systems are described, such as evanescent wave fluorescence and bioluminescent optical fibre biosensors, as well as interferometric, ellipsometric and reflectometric interference spectroscopy and surface-enhanced Raman scattering biosensors. The optical biosensors discussed here allow the sensitive and selective detection of a wide range of analytes including viruses, toxins, drugs, antibodies, tumour biomarkers and tumour cells. PMID:27365039

  18. Immobilization of biorecognition molecules on O2 plasma-functionalized SWCNT electrodes for biosensors

    NASA Astrophysics Data System (ADS)

    Jin, Joon-Hyung; Kim, Joon Hyub; Lee, Jun-Yong; Min, Nam Ki

    2012-11-01

    Biointerfaces capable of biological recognition and specificity are very important for the development of carbon nanotube based biosensors. Here, we explore experimentally the effects of O2 plasma treatment on the biomolecule immobilization properties of single-walled carbon nanotube (SWCNT) electrodes for electrochemical biosensing. The SWCNT film was integrated into an electrochemical three-electrode system on a glass substrate and then treated with an O2 plasma to improve its electrochemical response. Glucose oxidases, antibodies, and deoxyribonucleic acids (DNAs) were covalently immobilized on the plasma-functionalized (pf) SWCNT working electrodes, and the electrochemical and bioelectrocatalytic properties of three biomolecular assemblies generated on the pf-SWCNT electrodes were investigated using cyclic voltammetry, square-wave voltammetry, and chronoamperometry. The pf-SWCNT films were found to provide electrochemical biosensing electrodes having high electroactivity and sensitivity for detecting glucoses, antigens, and DNA molecules.

  19. DNA Sensors with Diamond as a Promising Alternative Transducer Material

    PubMed Central

    Vermeeren, Veronique; Wenmackers, Sylvia; Wagner, Patrick; Michiels, Luc

    2009-01-01

    Bio-electronics is a scientific field coupling the achievements in biology with electronics to obtain higher sensitivity, specificity and speed. Biosensors have played a pivotal role, and many have become established in the clinical and scientific world. They need to be sensitive, specific, fast and cheap. Electrochemical biosensors are most frequently cited in literature, often in the context of DNA sensing and mutation analysis. However, many popular electrochemical transduction materials, such as silicon, are susceptible to hydrolysis, leading to loss of bioreceptor molecules from the surface. Hence, increased attention has been shifted towards diamond, which surpasses silicon on many levels. PMID:22346717

  20. Functionalization of optical nanotip arrays with an electrochemical microcantilever for multiplexed DNA detection.

    PubMed

    Descamps, Emeline; Duroure, Nathalie; Deiss, Frédérique; Leichlé, Thierry; Adam, Catherine; Mailley, Pascal; Aït-Ikhlef, Ali; Livache, Thierry; Nicu, Liviu; Sojic, Neso

    2013-08-01

    Optical nanotip arrays fabricated on etched fiber bundles were functionalized with DNA spots. Such unconventional substrates (3D and non-planar) are difficult to pattern with standard microfabrication techniques but, using an electrochemical cantilever, up to 400 spots were electrodeposited on the nanostructured optical surface in 5 min. This approach allows each spot to be addressed individually and multiplexed fluorescence detection is demonstrated. Finally, remote fluorescence detection was performed by imaging through the optical fiber bundle itself after hybridisation with the complementary sequence. PMID:23695411

  1. Electrochemical Investigation of Interaction between a Bifunctional Probe and GG Mismatch Duplex.

    PubMed

    Li, Jiao; He, Hanping; Peng, Xiaoqian; Huang, Min; Zhang, Xiuhua; Wang, Shengfu

    2015-01-01

    A bifunctional probe (FecNC), containing a recognition part and an electrochemical active center, was applied to electrochemical detection of GG mismatch duplexes. The preparation of gold electrodes modified by mismatch and complementatry duplexes was characterized by electrochemical impedance spectroscopy (EIS) and optimized for better detection in terms of self-assembly time, hybridization time, and incubation time. The interaction between FecNC and DNA duplexes modified on the surface of a gold electrode was explored by square wave voltammetry (SWV) and EIS. The results showed that the DNA duplexes with GG mismatch on the surface of a gold electrode was easily detected by the largest electrochemical signal of the bifunctional probe because of its selective binding to GG mismatches. The bifunctional probe could offer a simple, effective electrochemical detection of GG mismatches, and theoretical bases for development of electrochemical biosensors. Further, the method would be favorable for diagnosis of genetic diseases. PMID:26165289

  2. Recent Trends in Biosensors

    NASA Astrophysics Data System (ADS)

    Karube, Isao

    The determination of organic compounds in foods is very important in food industries. A various compounds are contained in foods, selective determination methods are required for food processing and analysis. Electrochemical monitoring devices (biosensors) employing immobilized biocatalysts such as immobilized enzymes, organelles, microorganisms, and tissue have definite advantages. The enzyme Sensors consisted of immobilized enzymes and electrochemical devices. Enzyme sensors could be used for the determination of sugars, amino acids, organic acids, alcohols, lipids, nucleic acid derivatives, etc.. Furthermore, a multifunctional biosensor for the determination of several compounds has been developed for food processing. On the other hand, microbial sensors consisted of immobilized microorganisms and electrodes have been used for industrial and environmental analysis. Microbial sensors were applied for the determination of sugars, organic acids, alcohols, amino acids, mutagens, me thane, ammonia, and BOD. Furthermore, micro-biosensors using immobilized biocatalysts and ion sensitive field effect transistor or microelectrodes prepared by silicon fabrication technologies have been developed for medical ap. plication and food processing. This review summarizes the design and application of biosensors.

  3. The new generation drug candidate molecules: Spectral, electrochemical, DNA-binding and anticancer activity properties

    NASA Astrophysics Data System (ADS)

    Gölcü, Ayşegül; Muslu, Harun; Kılıçaslan, Derya; Çeşme, Mustafa; Eren, Özge; Ataş, Fatma; Demirtaş, İbrahim

    2016-09-01

    The new generation drug candidate molecules [Cu(5-Fu)2Cl2H2O] (NGDCM1) and [Zn(5-Fu)2(CH3COO)2] (NGDCM2) were obtained from the reaction of copper(II) and zinc(II) salts with the anticancer drug 5-fluoracil (5-Fu). These compounds have been characterized by spectroscopic and analytical techniques. Thermal behavior of the compounds were also investigated. The electrochemical properties of the compounds have been investigated by cyclic voltammetry (CV) using glassy carbon electrode. The biological activity of the NGDCM1 and NGDCM2 has been evaluated by examining their ability to bind to fish sperm double strand DNA (FSdsDNA) with UV spectroscopy. UV studies of the interaction of the 5-Fu and metal derivatives with FSdsDNA have shown that these compounds can bind to FSdsDNA. The binding constants of the compounds with FSdsDNA have also been calculated. Thermal decomposition of the compounds lead to the formation of CuO and ZnO as final products. The effect of proliferation 5-Fu, NGDCM1 and NGDCM2 were examined on the HeLa cells using real-time cell analyzer with three different concentrations.

  4. Highly Disordered Array of Silicon Nanowires: an Effective and Scalable Approach for Performing and Flexible Electrochemical Biosensors.

    PubMed

    Maiolo, Luca; Polese, Davide; Pecora, Alessandro; Fortunato, Guglielmo; Shacham-Diamand, Yosi; Convertino, Annalisa

    2016-03-01

    The direct integration of disordered arranged and randomly oriented silicon nanowires (SiNWs) into ultraflexible and transferable electronic circuits for electrochemical biosensing applications is proposed. The working electrode (WE) of a three-electrode impedance device, fabricated on a polyimide (PI) film, is modified with SiNWs covered by a thin Au layer and functionalized to bind the sensing element. The biosensing behavior is investigated through the ligand-receptor binding of biotin-avidin system. Impedance measurements show a very efficient detection of the avidin over a broad range of concentrations from hundreds of micromolar down to the picomolar values. The impedance response is modeled through a simple equivalent circuit, which takes into account the unique WE morphology and its modification with successive layers of biomolecules. This approach of exploiting highly disordered SiNW ensemble in biosensing proves to be very promising for the following three main reasons: first, the system morphology allows high sensing performance; second, these nanostructures can be built via scalable and transferable fabrication methodology allowing an easy integration on non-conventional substrates; third, reliable modeling of the sensing response can be developed by considering the morphological and surface characteristics over an ensemble of disordered NWs rather than over individual NWs. PMID:26717420

  5. Electrochemical biosensor based on glucose oxidase encapsulated within enzymatically synthesized poly(1,10-phenanthroline-5,6-dione).

    PubMed

    Ciftci, Hakan; Oztekin, Yasemin; Tamer, Ugur; Ramanaviciene, Almira; Ramanavicius, Arunas

    2014-11-01

    This study is focused on the investigation of electrocatalytic effect of glucose oxidase (GOx) immobilized on the graphite rod (GR) electrode. The enzyme modified electrode was prepared by encapsulation of immobilized GOx within enzymatically formed poly(1,10-phenanthroline-5,6-dione) (pPD) film. The electrochemical responses of such enzymatic electrode (pPD/GOx/GR) vs. different glucose concentrations were examined chronoamperometrically in acetate-phosphate buffer solution (A-PBS), pH 6.0, under aerobic or anaerobic conditions. Amperometric signals of the pPD/GOx/GR electrode exhibited well-defined hyperbolic dependence upon glucose concentration. Amperometric signals at 100mM of glucose were 41.17 and 32.27 μA under aerobic and anaerobic conditions, respectively. Amperometric signals of the pPD/GOx/GR electrode decreased by 6% within seven days. The pPD/GOx/GR electrode showed excellent selectivity in the presence of dopamine and uric acid. Furthermore it had a good reproducibility and repeatability with standard deviation of 9.4% and 8.0%, respectively. PMID:25454754

  6. Graphene, carbon nanotubes, zinc oxide and gold as elite nanomaterials for fabrication of biosensors for healthcare.

    PubMed

    Kumar, Sandeep; Ahlawat, Wandit; Kumar, Rajesh; Dilbaghi, Neeraj

    2015-08-15

    Technological advancements worldwide at rapid pace in the area of materials science and nanotechnology have made it possible to synthesize nanoparticles with desirable properties not exhibited by the bulk material. Among variety of available nanomaterials, graphene, carbon nanotubes, zinc oxide and gold nanopartilces proved to be elite and offered amazing electrochemical biosensing. This encourages us to write a review which highlights the recent achievements in the construction of genosensor, immunosensor and enzymatic biosensor based on the above nanomaterials. Carbon based nanomaterials offers a direct electron transfer between the functionalized nanomaterials and active site of bioreceptor without involvement of any mediator which not only amplifies the signal but also provide label free sensing. Gold shows affinity towards immunological molecules and is most routinely used for immunological sensing. Zinc oxide can easily immobilize proteins and hence offers a large group of enzyme based biosensor. Modification of the working electrode by introduction of these nanomaterials or combination of two/three of above nanomaterials together and forming a nanocomposite reflected the best results with excellent stability, reproducibility and enhanced sensitivity. Highly attractive electrochemical properties and electrocatalytic activity of these elite nanomaterials have facilitated achievement of enhanced signal amplification needed for the construction of ultrasensitive electrochemical affinity biosensors for detection of glucose, cholesterol, Escherichia coli, influenza virus, cancer, human papillomavirus, dopamine, glutamic acid, IgG, IgE, uric acid, ascorbic acid, acetlycholine, cortisol, cytosome, sequence specific DNA and amino acids. Recent researches for bedside biosensors are also discussed. PMID:25899923

  7. Triplex DNA: A new platform for polymerase chain reaction – based biosensor

    PubMed Central

    Li, Yubin; Miao, Xiangmin; Ling, Liansheng

    2015-01-01

    Non - specific PCR amplification and DNA contamination usually accompany with PCR process, to overcome these problems, here we establish a sensor for thrombin by sequence - specific recognition of the PCR product with molecular beacon through triplex formation. Probe A and probe B were designed for the sensor, upon addition of thrombin, two probes hybridized to each other and the probe B was extended in the presence of Klenow Fragment polymerase and dNTPs. The PCR amplification occurred with further addition of Taq DNA Polymerase and two primers, the PCR product was recognized by molecular beacon through triplex formation. The fluorescence intensity increased with the logarithm of the concentration of thrombin over the range from 1.0 × 10−12 M to 1.0 × 10−7 M, with a detection limit of 261 fM. Moreover, the effect of DNA contamination and non - specific amplification could be ignored completely in the proposed strategy. PMID:26268575

  8. Triplex DNA: A new platform for polymerase chain reaction-based biosensor.

    PubMed

    Li, Yubin; Miao, Xiangmin; Ling, Liansheng

    2015-01-01

    Non-specific PCR amplification and DNA contamination usually accompany with PCR process, to overcome these problems, here we establish a sensor for thrombin by sequence-specific recognition of the PCR product with molecular beacon through triplex formation. Probe A and probe B were designed for the sensor, upon addition of thrombin, two probes hybridized to each other and the probe B was extended in the presence of Klenow Fragment polymerase and dNTPs. The PCR amplification occurred with further addition of Taq DNA Polymerase and two primers, the PCR product was recognized by molecular beacon through triplex formation. The fluorescence intensity increased with the logarithm of the concentration of thrombin over the range from 1.0 × 10(-12) M to 1.0 × 10(-7) M, with a detection limit of 261 fM. Moreover, the effect of DNA contamination and non - specific amplification could be ignored completely in the proposed strategy. PMID:26268575

  9. The electrophotonic silicon biosensor.

    PubMed

    Juan-Colás, José; Parkin, Alison; Dunn, Katherine E; Scullion, Mark G; Krauss, Thomas F; Johnson, Steven D

    2016-01-01

    The emergence of personalized and stratified medicine requires label-free, low-cost diagnostic technology capable of monitoring multiple disease biomarkers in parallel. Silicon photonic biosensors combine high-sensitivity analysis with scalable, low-cost manufacturing, but they tend to measure only a single biomarker and provide no information about their (bio)chemical activity. Here we introduce an electrochemical silicon photonic sensor capable of highly sensitive and multiparameter profiling of biomarkers. Our electrophotonic technology consists of microring resonators optimally n-doped to support high Q resonances alongside electrochemical processes in situ. The inclusion of electrochemical control enables site-selective immobilization of different biomolecules on individual microrings within a sensor array. The combination of photonic and electrochemical characterization also provides additional quantitative information and unique insight into chemical reactivity that is unavailable with photonic detection alone. By exploiting both the photonic and the electrical properties of silicon, the sensor opens new modalities for sensing on the microscale. PMID:27624590

  10. DNA-based hybridization chain reaction for an ultrasensitive cancer marker EBNA-1 electrochemical immunosensor.

    PubMed

    Song, Chao; Xie, Guoming; Wang, Li; Liu, Lingzhi; Tian, Guang; Xiang, Hua

    2014-08-15

    An ultrasensitive and selective electrochemical immunosensor was developed for the detection of Epstein Barr virus nuclear antigen 1 (EBNA-1). Firstly, a suspension of graphene sheets (GS) and multi-walled carbon nanotubes (MWCNTs) was prepared with the aid of chitosan (CS) solution and then modified on a glassy carbon electrode (GCE). Gold nanoparticles (AuNPs) were then electrodeposited onto the surface of the GS-MWCNTs film by cyclic voltammetry (CV) to immobilize the captured antibodies. After that, specific sandwich immunoreactions were formed among the captured antibody, EBNA-1, and secondary antibody, DNA-coated carboxyl multi-wall carbon nanotubes (DNA-MWCNTs-Ab2). DNA initiator strands (S0) and secondary antibodies linked to the MWCNTs and double-helix DNA polymers were obtained by hybridization chain reaction (HCR), and here S0 on the MWCNTs propagates a chain reaction of hybridization events between two alternating hairpins to form a nicked double-helix. Finally, electroactive indicator doxorubicin hydrochloride was intercalated into the CG-GC steps between the HCR products and could produce an electrochemical signal, which was monitored by differential pulse voltammetry (DPV). Under optimum conditions, the amperometric signal increased linearly with the target concentrations (0.05-6.4ngmL(-1)), and the immunosensor exhibited a detection limit as low as 0.7pgmL(-1) (S/N=3). The proposed method showed acceptable stability and reproducibility, as well as favorable recovery for EBNA-1 in human serum. The proposed immunosensor provides a novel avenue for signal amplification and potential applications in bioanalysis and clinical diagnostics. PMID:24632131

  11. Photoinduced interactions of supramolecular ruthenium(II) complexes with plasmid DNA: synthesis and spectroscopic, electrochemical, and DNA photocleavage studies.

    PubMed

    Swavey, Shawn; DeBeer, Madeleine; Li, Kaiyu

    2015-04-01

    Two new bridging ligands have been synthesized by combining substituted benzaldehydes with phenanthrolinopyrrole (php), resulting in new polyazine bridging ligands. The ligands have been characterized by (1)H NMR, mass spectroscopy, and elemental analysis. These new ligands display π-π* transitions above 500 nm with modest molar absorptivities. Upon excitation at the ligand-centered charge-transfer transition, weak emission with a maximum wavelength of 612 nm is observed. When coordinated to two ruthenium(II) bis(bipyridyl) groups, the new bimetallic complexes generated give an overall 4+ charge. The electronic transitions of the bimetallic ruthenium(II) complexes display traditional π-π* transitions at 287 nm and metal-to-ligand charge-transfer transitions at 452 nm with molar absorptivities greater than 30000 M(-1) cm(-1). Oxidation of the ruthenium(II) metal centers to ruthenium(III) occurs at potentials above 1.4 V versus the Ag/AgCl reference electrode. Spectroscopic and electrochemical measurements indicate that the ruthenium(II) moieties behave independently. Both complexes are water-soluble and show the ability to photonick plasmid DNA when irradiated with low-energy light above 550 nm. In addition, one of the complexes, [Ru(bpy)2php]2Van(4+), shows the ability to linearize plasmid DNA and gives evidence, by gel electrophoresis, of photoinduced binding to plasmid DNA. PMID:25798576

  12. Sensitive and direct electrochemical detection of double-stranded DNA utilizing alkaline phosphatase-labelled zinc finger proteins.

    PubMed

    Noh, Soodong; Ha, Dat Thinh; Yang, Haesik; Kim, Moon-Soo

    2015-06-21

    Direct detection of double-stranded DNA (dsDNA) using zinc finger proteins (ZFPs) is of great importance in biomedical applications such as identifying pathogens and circulating DNAs. However, its sensitivity is still not sufficiently high because limited signalling labels can be conjugated or fused. Herein, we report sensitive and direct detection of dsDNA using (i) alkaline phosphatase (ALP) as a fast catalytic label conjugated to ZFPs along with (ii) electrochemical measurement of an ALP product (l-ascorbic acid) at the indium-tin oxide electrode with a high signal-to-background ratio. ALP is simply conjugated to a ZFP through lysine residues in a ZFP purification tag, a maltose binding protein (MBP). Sandwich-type electrochemical detection of dsDNA allows a detection limit of ca. 100 fM without using DNA amplification. PMID:25969923

  13. Functionalized Solid Electrodes for Electrochemical Biosensing of Purine Nucleobases and Their Analogues: A Review

    PubMed Central

    Sharma, Vimal Kumar; Jelen, Frantisek; Trnkova, Libuse

    2015-01-01

    Interest in electrochemical analysis of purine nucleobases and few other important purine derivatives has been growing rapidly. Over the period of the past decade, the design of electrochemical biosensors has been focused on achieving high sensitivity and efficiency. The range of existing electrochemical methods with carbon electrode displays the highest rate in the development of biosensors. Moreover, modification of electrode surfaces based on nanomaterials is frequently used due to their extraordinary conductivity and surface to volume ratio. Different strategies for modifying electrode surfaces facilitate electron transport between the electrode surface and biomolecules, including DNA, oligonucleotides and their components. This review aims to summarize recent developments in the electrochemical analysis of purine derivatives, as well as discuss different applications. PMID:25594595

  14. Quantum-dot biosensor for hybridization and detection of R3500Q mutation of apolipoprotein B-100 gene.

    PubMed

    Mazloum-Ardakani, Mohammad; Aghaei, Roghayyeh; Heidari, Mohammad Mehdi

    2015-10-15

    A quantum-dot electrode system was developed as a transducer surface for covalent immobilization of a designed synthetic ApoB-100 specific probe, DNA hybridization and monitoring of DNA synthesis for the sensitive detection of R3500Q mutation of apolipoprotein B-100 (ApoB-100) gene. CdS-QDs cause an improvement in the fundamental characteristics of the electrode interface, such as its electroactive surface area, diffusion coefficient and electron transfer kinetics. The sensing characteristics of this biosensor offer a suitable potential for detection of target oligonucleotide with a detection limit of 3.4 × 10(-17)M. Also, the electrochemical responses of single-stranded DNA (ssDNA), DNA hybridization and DNA synthesis were investigated using electrochemical impedance spectroscopy (EIS). The extracted genomic DNA was detected based on changes in the charge transfer resistance (RCT) with [Fe(CN)6](3-/4-) as a redox probe. The proposed biosensor can distinguish between the normal sequence and the mutant sequence of ApoB-100 gene, promising a possibility to apply the QD-based biosensor for clinical investigations. PMID:26022781

  15. A novel quantum dot nanocluster as versatile probe for electrochemiluminescence and electrochemical assays of DNA and cancer cells.

    PubMed

    Jie, Guifen; Zhang, Jian; Jie, Guixia; Wang, Lei

    2014-02-15

    A novel dendritic quantum dot (QD) nanocluster was constructed and used as versatile electrochemiluminescence (ECL) and electrochemical probe for the detection of DNA and cancer cells. Owing to the many functional groups present in the nanoclusters, a large number of QDs were assembled on the nanoclusters, which could greatly amplify both the ECL and electrochemical signals of QDs. Carbon nanotubes (CNTs)/gold nanoparticles' (NPs) hybrids were used as amplified platform for assembling large numbers of DNA on the electrode, which also improve the bioactivity and stability of the electrode. After the QD-DNA signal probe was recognized with target DNA (t-DNA), the amplified ECL signal for the detection of target DNA was obtained. Furthermore, magnetic nanoparticles were employed for cell aptamers immobilization, the same QD nanocluster-DNA probe was also extended for electrochemical detection of cancer cells using sensitive anodic stripping voltammetry (ASV) method, which simplified the separation procedures and improved the sensitivity. It is anticipated that the assays could provide promising and cost effective approach for the early and accurate detection of DNA and cancer cells. PMID:24021658

  16. Immobilization of Enzymes by Electrochemical and Chemical Oxidative Polymerization of L-DOPA to Fabricate Amperometric Biosensors and Biofuel Cells.

    PubMed

    Dai, Mengzhen; Sun, Lingen; Chao, Long; Tan, Yueming; Fu, Yingchun; Chen, Chao; Xie, Qingji

    2015-05-27

    Electrochemical/chemical oxidative synthesis and biosensing/biofuel cell applications of poly(L-DOPA) (PD) are studied versus polydopamine (PDA) as a recent hotspot biomaterial. The enzyme electrode developed by coelectrodeposition of PD and glucose oxidase (GOx), uricase, or tyrosinase shows biosensing performance superior to that of the corresponding PDA-based enzyme electrode. The chemical oxidative polymerization of L-DOPA (PDC) by NaAuCl4 in GOx-containing neutral aqueous solution is used to immobilize GOx and gold nanoparticles (AuNPs). The thus-prepared chitosan (CS)/GOx-PDC-AuNPs/Au(plate)/Au electrode working in the first-generation biosensing mode responds linearly to glucose concentration with a sensitivity of 152 μA mM(-1) cm(-2), which is larger than those of the CS/GOx-PDAC-AuNPs/Au(plate)/Au electrode, the CS/GOx-poly(3-anilineboronic acid) (PABA)-AuNPs/Au(plate)/Au electrode, and the most reported GOx-based enzyme electrodes. This PDC-based enzyme electrode also works well in the second-generation biosensing mode and as an excellent bioanode in biofuel cell construction, probably because PD as an amino acid polymer has the higher biocompatibility and the more favorable affinity to the enzyme than PDA. The PD material of great convenience in synthesis, outstanding biocompatibility for preparing high-performance bionanocomposites, and strong capability of multifunctional coatings on many surfaces may find wide applications in diversified fields including biotechnology and surface-coating. PMID:25938891

  17. G-quadruplex DNA biosensor for sensitive visible detection of genetically modified food.

    PubMed

    Jiang, Xiaohua; Zhang, Huimin; Wu, Jun; Yang, Xiang; Shao, Jingwei; Lu, Yujing; Qiu, Bin; Lin, Zhenyu; Chen, Guonan

    2014-10-01

    In this paper, a novel label-free G-quadruplex DNAzyme sensor has been proposed for colorimetric identification of GMO using CaMV 35S promoter sequence as the target. The binary probes can fold into G-quadruplex structure in the presence of DNA-T (Target DNA) and then combine with hemin to form a DNAzyme resembling horseradish peroxidase. The detection system consists of two G-rich probes with 2:2 split mode by using the absorbance and color of ABTS(2-) as signal reporter. Upon the addition of a target sequence, two probes both hybridize with target and then their G-rich sequences combine to form a G-quadruplex DNAzyme, and the DNAzyme can catalyze the reaction of ABTS(2-) with H2O2. Then the linear range is from 0.05 to 0.5 μM while detection limit is 5nM. These results demonstrate that the proposed G-quadruplex DNAzyme method could be used as a simple, sensitive and cost-effective approach for assays of GMO. PMID:25059184

  18. Using triplex-forming oligonucleotide probes for the reagentless, electrochemical detection of double-stranded DNA.

    PubMed

    Patterson, Adriana; Caprio, Felice; Vallée-Bélisle, Alexis; Moscone, Danila; Plaxco, Kevin W; Palleschi, Giuseppe; Ricci, Francesco

    2010-11-01

    We report a reagentless, electrochemical sensor for the detection of double-stranded DNA targets that employs triplex-forming oligonucleotides (TFOs) as its recognition element. These sensors are based on redox-tagged TFO probes strongly chemisorbed onto an interrogating gold electrode. Upon the addition of the relevant double-stranded DNA target, the probe forms a rigid triplex structure via reverse Hoogsteen base pairing in the major groove. The formation of the triplex impedes contact between the probe's redox moiety and the interrogati