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Sample records for dna biosensor built

  1. 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

  2. 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

  3. 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.

  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. 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.

  6. 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...

  7. 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

  8. 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.

  9. 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

  10. 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.

  11. 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

  12. 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

  13. 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

  14. 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

  15. Bienzyme biosensors for glucose, ethanol and putrescine built on oxidase and sweet potato peroxidase.

    PubMed

    Castillo, Jaime; Gáspár, Szilveszter; Sakharov, Ivan; Csöregi, Elisabeth

    2003-05-01

    Amperometric biosensors for glucose, ethanol, and biogenic amines (putrescine) were constructed using oxidase/peroxidase bienzyme systems. The H(2)O(2) produced by the oxidase in reaction with its substrate is converted into a measurable signal via a novel peroxidase purified from sweet potato peels. All developed biosensors are based on redox hydrogels formed of oxidases (glucose oxidase, alcohol oxidase, or amine oxidase) and the newly purified sweet potato peroxidase (SPP) cross-linked to a redox polymer. The developed electrodes were characterized (sensitivity, stability, and performances in organic medium) and compared with similarly built ones using the 'classical' horseradish peroxidase (HRP). The SPP-based electrodes displayed higher sensitivity and better detection limit for putrescine than those using HRP and were also shown to retain their activity in organic phase much better than the HPR based ones. The importance of attractive or repulsive electrostatic interactions between the peroxidases and oxidases (determined by their isoelectric points) were found to play an important role in the sensitivity of the obtained sensors. PMID:12706582

  16. 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

  17. 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.

  18. 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

  19. 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

  20. 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

  1. 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

  2. Digital biosensors with built-in logic for biomedical applications--biosensors based on a biocomputing concept.

    PubMed

    Wang, Joseph; Katz, Evgeny

    2010-10-01

    This article reviews biomolecular logic systems for bioanalytical applications, specifically concentrating on the prospects and fundamental and practical challenges of designing digitally operating biosensors logically processing multiple biochemical signals. Such digitally processed information produces a final output in the form of a yes/no response through Boolean logic networks composed of biomolecular systems, and hence leads to a high-fidelity biosensing compared with traditional single (or parallel) sensing devices. It also allows direct coupling of the signal processing with chemical actuators to produce integrated "smart" "sense/act" (biosensor-bioactuator) systems. Unlike common biosensing devices based on a single input (analyte), devices based on biochemical logic systems require a fundamentally new approach for the sensor design and operation and careful attention to the interface of biocomputing systems and electronic transducers. As common in conventional biosensors, the success of the enzyme logic biosensor would depend, in part, on the immobilization of the biocomputing reagent layer. Such surface confinement provides a contact between the biocomputing layer and the transducing surface and combines efficiently the individual logic-gate elements. Particular attention should thus be given to the composition, preparation, and immobilization of the biocomputing surface layer, to the role of the system scalability, and to the efficient transduction of the output signals. By processing complex patterns of multiple physiological markers, such multisignal digital biosensors should have a profound impact upon the rapid diagnosis and treatment of diseases, and particularly upon the timely detection and alert of medical emergencies (along with immediate therapeutic intervention). Other fields ranging from biotechnology to homeland security would benefit from these advances in new biocomputing biosensors and the corresponding closed-loop "add/act" operation

  3. 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...

  4. 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

  5. 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.

  6. 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

  7. 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

  8. 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

  9. 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.

  10. 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

  11. 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.

  12. 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)

  13. "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

  14. 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

  15. 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.

  16. 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'.

  17. 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

  18. 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).

  19. Biosensors with Built-In Biomolecular Logic Gates for Practical Applications

    PubMed Central

    Lai, Yu-Hsuan; Sun, Sin-Cih; Chuang, Min-Chieh

    2014-01-01

    Molecular logic gates, designs constructed with biological and chemical molecules, have emerged as an alternative computing approach to silicon-based logic operations. These molecular computers are capable of receiving and integrating multiple stimuli of biochemical significance to generate a definitive output, opening a new research avenue to advanced diagnostics and therapeutics which demand handling of complex factors and precise control. In molecularly gated devices, Boolean logic computations can be activated by specific inputs and accurately processed via bio-recognition, bio-catalysis, and selective chemical reactions. In this review, we survey recent advances of the molecular logic approaches to practical applications of biosensors, including designs constructed with proteins, enzymes, nucleic acids, nanomaterials, and organic compounds, as well as the research avenues for future development of digitally operating “sense and act” schemes that logically process biochemical signals through networked circuits to implement intelligent control systems. PMID:25587423

  20. Biosensors with built-in biomolecular logic gates for practical applications.

    PubMed

    Lai, Yu-Hsuan; Sun, Sin-Cih; Chuang, Min-Chieh

    2014-09-01

    Molecular logic gates, designs constructed with biological and chemical molecules, have emerged as an alternative computing approach to silicon-based logic operations. These molecular computers are capable of receiving and integrating multiple stimuli of biochemical significance to generate a definitive output, opening a new research avenue to advanced diagnostics and therapeutics which demand handling of complex factors and precise control. In molecularly gated devices, Boolean logic computations can be activated by specific inputs and accurately processed via bio-recognition, bio-catalysis, and selective chemical reactions. In this review, we survey recent advances of the molecular logic approaches to practical applications of biosensors, including designs constructed with proteins, enzymes, nucleic acids, nanomaterials, and organic compounds, as well as the research avenues for future development of digitally operating "sense and act" schemes that logically process biochemical signals through networked circuits to implement intelligent control systems. PMID:25587423

  1. 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

  2. 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.

  3. 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.

  4. 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

  5. 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

  6. 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

  7. Ancient whole genome enrichment using baits built from modern DNA.

    PubMed

    Enk, Jacob M; Devault, Alison M; Kuch, Melanie; Murgha, Yusuf E; Rouillard, Jean-Marie; Poinar, Hendrik N

    2014-05-01

    We report metrics from complete genome capture of nuclear DNA from extinct mammoths using biotinylated RNAs transcribed from an Asian elephant DNA extract. Enrichment of the nuclear genome ranged from 1.06- to 18.65-fold, to an apparent maximum threshold of ∼80% on-target. This projects an order of magnitude less costly complete genome sequencing from long-dead organisms, even when a reference genome is unavailable for bait design. PMID:24531081

  8. 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

  9. 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.

  10. 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...

  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. 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

  13. 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

  14. 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

  15. 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

  16. 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.

  17. 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

  18. 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

  19. 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

  20. 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

  1. 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

  2. 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

  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. 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

  5. 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

  6. 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.

  7. [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

  8. 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.

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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

  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. 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

  2. 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.

  3. 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

  4. 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

  5. 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

  6. 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

  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. 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.

  9. 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

  10. 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.

  11. 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

  12. 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

  13. 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

  14. 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.

  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. 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

  17. 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

  18. 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.

  19. 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.

  20. 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.

  1. 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

  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. 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

  4. 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.

  5. 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.

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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.

  12. 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

  13. 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

  14. 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

  15. 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.

  16. 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.

  17. 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

  18. 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

  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 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

  1. 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

  2. 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.

  3. 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.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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

  9. Prescribed nanoparticle cluster architectures and low-dimensional arrays built using octahedral DNA origami frames.

    PubMed

    Tian, Ye; Wang, Tong; Liu, Wenyan; Xin, Huolin L; Li, Huilin; Ke, Yonggang; Shih, William M; Gang, Oleg

    2015-07-01

    Three-dimensional mesoscale clusters that are formed from nanoparticles spatially arranged in pre-determined positions can be thought of as mesoscale analogues of molecules. These nanoparticle architectures could offer tailored properties due to collective effects, but developing a general platform for fabricating such clusters is a significant challenge. Here, we report a strategy for assembling three-dimensional nanoparticle clusters that uses a molecular frame designed with encoded vertices for particle placement. The frame is a DNA origami octahedron and can be used to fabricate clusters with various symmetries and particle compositions. Cryo-electron microscopy is used to uncover the structure of the DNA frame and to reveal that the nanoparticles are spatially coordinated in the prescribed manner. We show that the DNA frame and one set of nanoparticles can be used to create nanoclusters with different chiroptical activities. We also show that the octahedra can serve as programmable interparticle linkers, allowing one- and two-dimensional arrays to be assembled with designed particle arrangements. PMID:26005999

  10. Prescribed nanoparticle cluster architectures and low-dimensional arrays built using octahedral DNA origami frames

    DOE PAGESBeta

    Tian, Ye; Wang, Tong; Liu, Wenyan; Xin, Huolin L.; Li, Huilin; Ke, Yonggang; Shih, William M.; Gang, Oleg

    2015-05-25

    Three-dimensional mesoscale clusters that are formed from nanoparticles spatially arranged in pre-determined positions can be thought of as mesoscale analogues of molecules. These nanoparticle architectures could offer tailored properties due to collective effects, but developing a general platform for fabricating such clusters is a significant challenge. Here, we report a strategy for assembling 3D nanoparticle clusters that uses a molecular frame designed with encoded vertices for particle placement. The frame is a DNA origami octahedron and can be used to fabricate clusters with various symmetries and particle compositions. Cryo-electron microscopy is used to uncover the structure of the DNA framemore » and to reveal that the nanoparticles are spatially coordinated in the prescribed manner. We show that the DNA frame and one set of nanoparticles can be used to create nanoclusters with different chiroptical activities. We also show that the octahedra can serve as programmable interparticle linkers, allowing one- and two-dimensional arrays to be assembled that have designed particle arrangements.« less

  11. Prescribed nanoparticle cluster architectures and low-dimensional arrays built using octahedral DNA origami frames

    SciTech Connect

    Tian, Ye; Wang, Tong; Liu, Wenyan; Xin, Huolin L.; Li, Huilin; Ke, Yonggang; Shih, William M.; Gang, Oleg

    2015-05-25

    Three-dimensional mesoscale clusters that are formed from nanoparticles spatially arranged in pre-determined positions can be thought of as mesoscale analogues of molecules. These nanoparticle architectures could offer tailored properties due to collective effects, but developing a general platform for fabricating such clusters is a significant challenge. Here, we report a strategy for assembling 3D nanoparticle clusters that uses a molecular frame designed with encoded vertices for particle placement. The frame is a DNA origami octahedron and can be used to fabricate clusters with various symmetries and particle compositions. Cryo-electron microscopy is used to uncover the structure of the DNA frame and to reveal that the nanoparticles are spatially coordinated in the prescribed manner. We show that the DNA frame and one set of nanoparticles can be used to create nanoclusters with different chiroptical activities. We also show that the octahedra can serve as programmable interparticle linkers, allowing one- and two-dimensional arrays to be assembled that have designed particle arrangements.

  12. 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.

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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.

  1. 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

  2. [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

  3. 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

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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

  12. 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.

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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.

  1. 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.

  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. 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

  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. 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

  6. 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.

  7. 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.

  8. 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

  9. 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

  10. 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

  11. 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

  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. 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

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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

  1. Label-free fluorescent biosensor based on the target recycling and Thioflavin T-induced quadruplex formation for short DNA species of c-erbB-2 detection.

    PubMed

    Chen, Jinghua; Lin, Jia; Zhang, Xi; Cai, Shuxian; Wu, Dongzhi; Li, Chunyan; Yang, Sheng; Zhang, Jing

    2014-03-19

    Non-invasive early diagnosis of breast cancer is the most effective way to improve the survival rate and increase more chances of breast-conserving. In this paper, we developed a label-free fluorescent biosensor based on nuclease assisted target recycling and Thioflavin T-induced quadruplex formation for short DNA species of c-erbB-2 detection in saliva. By employing the strategy, the sensor can detect as low as 20fM target DNA with high discrimination ability even against single-base mismatch sequence. To the best of our knowledge, the proposed sensor is the first attempt to apply Thioflavin T that possesses outstanding structural selectivity for G-quadruplex in DNA amplification techniques, which may represent a promising path toward direct breast cancer detection in saliva at the point of care. PMID:24594816

  2. A FRET-Based DNA Biosensor Tracks OmpR-Dependent Acidification of Salmonella during Macrophage Infection

    PubMed Central

    Chakraborty, Smarajit; Mizusaki, Hideaki; Kenney, Linda J.

    2015-01-01

    In bacteria, one paradigm for signal transduction is the two-component regulatory system, consisting of a sensor kinase (usually a membrane protein) and a response regulator (usually a DNA binding protein). The EnvZ/OmpR two-component system responds to osmotic stress and regulates expression of outer membrane proteins. In Salmonella, EnvZ/OmpR also controls expression of another two-component system SsrA/B, which is located on Salmonella Pathogenicity Island (SPI) 2. SPI-2 encodes a type III secretion system, which functions as a nanomachine to inject bacterial effector proteins into eukaryotic cells. During the intracellular phase of infection, Salmonella switches from assembling type III secretion system structural components to secreting effectors into the macrophage cytoplasm, enabling Salmonella to replicate in the phagocytic vacuole. Major questions remain regarding how bacteria survive the acidified vacuole and how acidification affects bacterial secretion. We previously reported that EnvZ sensed cytoplasmic signals rather than extracellular ones, as intracellular osmolytes altered the dynamics of a 17-amino-acid region flanking the phosphorylated histidine. We reasoned that the Salmonella cytoplasm might acidify in the macrophage vacuole to activate OmpR-dependent transcription of SPI-2 genes. To address these questions, we employed a DNA-based FRET biosensor (“I-switch”) to measure bacterial cytoplasmic pH and immunofluorescence to monitor effector secretion during infection. Surprisingly, we observed a rapid drop in bacterial cytoplasmic pH upon phagocytosis that was not predicted by current models. Cytoplasmic acidification was completely dependent on the OmpR response regulator, but did not require known OmpR-regulated genes such as ompC, ompF, or ssaC (SPI-2). Microarray analysis highlighted the cadC/BA operon, and additional experiments confirmed that it was repressed by OmpR. Acidification was blocked in the ompR null background in a Cad

  3. Time-resolved luminescence biosensor for continuous activity detection of protein acetylation-related enzymes based on DNA-sensitized terbium(III) probes.

    PubMed

    Han, Yitao; Li, Hao; Hu, Yufang; Li, Pei; Wang, Huixia; Nie, Zhou; Yao, Shouzhuo

    2015-09-15

    Protein acetylation of histone is an essential post-translational modification (PTM) mechanism in epigenetic gene regulation, and its status is reversibly controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). Herein, we have developed a sensitive and label-free time-resolved luminescence (TRL) biosensor for continuous detection of enzymatic activity of HATs and HDACs, respectively, based on acetylation-mediated peptide/DNA interaction and Tb(3+)/DNA luminescent probes. Using guanine (G)-rich DNA-sensitized Tb(3+) luminescence as the output signal, the polycationic substrate peptides interact with DNA with high affinity and subsequently replace Tb(3+), eliminating the luminescent signal. HAT-catalyzed acetylation remarkably reduces the positive charge of the peptides and diminishes the peptide/DNA interaction, resulting in the signal on detection via recovery of DNA-sensitized Tb(3+) luminescence. With this TRL sensor, HAT (p300) can be sensitively detected with a wide linear range from 0.2 to 100 nM and a low detection limit of 0.05 nM. The proposed sensor was further used to continuously monitor the HAT activity in real time. Additionally, the TRL biosensor was successfully applied to evaluating HAT inhibition by two specific inhibitors, anacardic acid and C464, and satisfactory Z'-factors above 0.73 were obtained. Moreover, this sensor is feasible to continuously monitor the HDAC (Sirt1)-catalyzed deacetylation with a linear range from 0.5 to 500 nM and a detection limit of 0.5 nM. The proposed sensor is a convenient, sensitive, and mix-and-read assay, presenting a promising platform for protein acetylation-targeted epigenetic research and drug discovery. PMID:26307596

  4. Approaches to label-free flexible DNA biosensors using low-temperature solution-processed InZnO thin-film transistors.

    PubMed

    Jung, Joohye; Kim, Si Joon; Lee, Keun Woo; Yoon, Doo Hyun; Kim, Yeong-Gyu; Kwak, Hee Young; Dugasani, Sreekantha Reddy; Park, Sung Ha; Kim, Hyun Jae

    2014-05-15

    Low-temperature solution-processed In-Zn-O (IZO) thin-film transistors (TFTs) exhibiting a favorable microenvironment for electron transfer by adsorbed artificial deoxyribonucleic acid (DNA) have extraordinary potential for emerging flexible biosensor applications. Superb sensing ability to differentiate even 0.5 μL of 50 nM DNA target solution was achieved through using IZO TFTs fabricated at 280 °C. Our IZO TFT had a turn-on voltage (V(on)) of -0.8 V, on/off ratio of 6.94 × 10(5), and on-current (I(on)) value of 2.32 × 10(-6)A in pristine condition. A dry-wet method was applied to immobilize two dimensional double crossover tile based DNA nanostructures on the IZO surface, after which we observed a negative shift of the transfer curve accompanied by a significant increase in the Ion and degradation of the Von and on/off ratio. As the concentration of DNA target solution increased, variances in these parameters became increasingly apparent. The sensing mechanism based on the current evolution was attributed to the oxidation of DNA, in which the guanine nucleobase plays a key role. The sensing behavior obtained from flexible biosensors on a polymeric substrate fabricated under the identical conditions was exactly analogous. These results compare favorably with the conventional field-effect transistor based DNA sensors by demonstrating remarkable sensitivity and feasibility of flexible devices that arose from a different sensing mechanism and a low-temperature process, respectively. PMID:24368226

  5. Selection and characterization of DNA aptamers for the development of light-up biosensor to detect Cd(II).

    PubMed

    Wang, Hongyan; Cheng, Hui; Wang, Jine; Xu, Lijun; Chen, Hongxia; Pei, Renjun

    2016-07-01

    In order to develop a facile, cost-effective and quick-testing light-up biosensor with excellent specificity for cadmium ions (Cd(II)) detection, a modified selection method based on target-induced release of strands was used to isolate aptamers of Cd (II) with high specificity. Circular Dichroism (CD) data confirmed that one of the selected aptamers underwent a distinct conformational change on addition of Cd (II). A biosensor for Cd(II) was developed based on the Cd(II)-induced release of fluorescence-labeled aptamer from complex with a quencher-labeled short complementary sequence. The sensing platform displayed a Cd(II) concentration-dependent increase of fluorescence intensity in the low micromolar range and had an excellent selectivity in the presence of various interfering metal ions. Such biosensor could potentially be used for the detection of Cd(II) in environmental samples. PMID:27154706

  6. A label-free and enzyme-free ultra-sensitive transcription factors biosensor using DNA-templated copper nanoparticles as fluorescent indicator and hairpin DNA cascade reaction as signal amplifier.

    PubMed

    Sha, Liang; Zhang, Xiaojun; Wang, Guangfeng

    2016-08-15

    Detection and quantification of specific protein with ultralow concentration play a crucial role in biotechnological applications and biomedical diagnostics. In this paper, a label-free and enzyme-free amplified fluorescent biosensor has been developed for transcription factors detection based on AT-rich double-stranded DNA-templated copper nanoparticles (ds DNA/Cu NPs) and hairpin DNA cascade reaction. This strategy was demonstrated by using nuclear factor-kappa B p50 (NF-κB p50) and specific recognition sequences as a model case. In this assay, a triplex consists of double-stranded DNA containing NF-κB p50 specifically binding sequences and a special design single-stranded DNA (Trigger) which is able to activate the hairpin DNA cascade amplifier (HDCA). In the presence of NF-κB p50, the triplex became unstable since the target bound to the recognition sequences with strong affinity. The selective binding event confirmed that the Trigger was successfully released from the triplex and initiated HDCA to yield the product which could effectively template the formation of fluorescent Cu NPs. The experimental results revealed that the advanced strategy was ultra-sensitive for detecting NF-κB p50 in the concentration range from 0.1 to 1000 pM with a detection limit of 0.096 pM. In addition, the relative standard deviation was 4.08% in 3 repetitive assays of 500 pM NF-κB p50, which indicated that the reproducibility of this strategy was acceptable. Besides desirable sensitivity, the developed biosensor also showed high selectivity, cost-effective, and simplified operations. In addition, the proposed biosensing platform is versatile. By conjugating with various specific recognition units, it could hold considerable potential to sensitive and selective detect various DNA-binding proteins and might find wide applications in biomedical fields. PMID:27045526

  7. Biosensors and their applications – A review

    PubMed Central

    Mehrotra, Parikha

    2016-01-01

    The various types of biosensors such as enzyme-based, tissue-based, immunosensors, DNA biosensors, thermal and piezoelectric biosensors have been deliberated here to highlight their indispensable applications in multitudinous fields. Some of the popular fields implementing the use of biosensors are food industry to keep a check on its quality and safety, to help distinguish between the natural and artificial; in the fermentation industry and in the saccharification process to detect precise glucose concentrations; in metabolic engineering to enable in vivo monitoring of cellular metabolism. Biosensors and their role in medical science including early stage detection of human interleukin-10 causing heart diseases, rapid detection of human papilloma virus, etc. are important aspects. Fluorescent biosensors play a vital role in drug discovery and in cancer. Biosensor applications are prevalent in the plant biology sector to find out the missing links required in metabolic processes. Other applications are involved in defence, clinical sector, and for marine applications. PMID:27195214

  8. Biosensors and their applications - A review.

    PubMed

    Mehrotra, Parikha

    2016-01-01

    The various types of biosensors such as enzyme-based, tissue-based, immunosensors, DNA biosensors, thermal and piezoelectric biosensors have been deliberated here to highlight their indispensable applications in multitudinous fields. Some of the popular fields implementing the use of biosensors are food industry to keep a check on its quality and safety, to help distinguish between the natural and artificial; in the fermentation industry and in the saccharification process to detect precise glucose concentrations; in metabolic engineering to enable in vivo monitoring of cellular metabolism. Biosensors and their role in medical science including early stage detection of human interleukin-10 causing heart diseases, rapid detection of human papilloma virus, etc. are important aspects. Fluorescent biosensors play a vital role in drug discovery and in cancer. Biosensor applications are prevalent in the plant biology sector to find out the missing links required in metabolic processes. Other applications are involved in defence, clinical sector, and for marine applications. PMID:27195214

  9. Detection of DNA utilizing a fluorescent reversible change of a biosensor based on the electron transfer from quantum dots to polymyxin B sulfate.

    PubMed

    Wang, Linlin; Liu, Shaopu; Liang, Wanjun; Li, Dan; Yang, Jidong; He, Youqiu

    2015-06-15

    A fluorescent "turn off-on" pattern for the detection of herring sperm DNA (hsDNA) had been designed through utilizing the interaction between polymyxin B sulfate (PMBS) and hsDNA as an inherent performance and the fluorescent transformation of glutathione (GSH)-capped CdTe quantum dots (QDs) as an external manifestation. Due to the occurrence of the photoinduced electron transfer from the QDs to PMBS, the fluorescence of GSH-capped CdTe QDs could be effectively quenched by PMBS, causing the system into "off" state. With the addition of hsDNA, the quenched fluorescence of GSH-capped CdTe QDs could be restored for the reason that PMBS embedded into hsDNA double helix structure to form new complex and peeled off from the surface of GSH-capped CdTe QDs, leading the system into "on" condition. Corresponding experimental results illustrated that the relative recovered fluorescence intensity of GSH-capped CdTe QDs-PMBS system was near proportional to the concentration of hsDNA within the range of 0.059-15.0 μg mL(-1). This proposed method demonstrated a good linear correlation coefficient of 0.9937 and a detection limit (3 σ/K) of 0.018 μg mL(-1) for hsDNA. This dual-directional fluorescent biosensor overcame the selectivity problem commonly existed in the traditional mono-directional fluorescence detection mode and owned perfect analysis applications in biochemical DNA monitoring. PMID:25744859

  10. Plasmonic nanoparticles: Towards the fabrication of biosensors

    NASA Astrophysics Data System (ADS)

    Shen, Hui

    2015-07-01

    Au and Ag nanoparticles are mainly employed in the fabrication of biosensors owing to their unique optical properties compared to other noble metal nanoparticles. Many biosensors are fabricated for the rapid detection of different analytes such as organic and inorganic molecules, biomolecules like DNA, proteins, biotoxins and pathogens. In this mini review we mainly discuss on the usage of Au and Ag nanoparticles for the fabrication of colorimetric, SERS and two photon based photoluminescence biosensors.

  11. Enhancing capacitive DNA biosensor performance by target overhang with application on screening test of HLA-B*58:01 and HLA-B*57:01 genes.

    PubMed

    Thipmanee, Orawan; Numnuam, Apon; Limbut, Warakorn; Buranachai, Chittanon; Kanatharana, Proespichaya; Vilaivan, Tirayut; Hirankarn, Nattiya; Thavarungkul, Panote

    2016-08-15

    A highly sensitive label-free DNA biosensor based on PNA probes immobilized on a gold electrode was used to detect a hybridization event. The effect of a target DNA overhang on the hybridization efficiency was shown to enhance the detected signal and allowed detection at a very low concentration. The sensors performances were investigated with a complementary target that had the same length as the probe, and the signal was compared to the target DNAs with different lengths and overhangs. A longer target DNA overhang was found to provide a better response. When the overhang was on the electrode side the signal enhancement was greater than when the overhang was on the solution side due to the increased thickness of the sensing surface, hence produced a larger capacitance change. Using conformationally constrained acpcPNA probes, double stranded DNA was detected sensitively and specifically without any denaturing step. When two acpcPNA probes were applied for the screening test for the double stranded HLA-B*58:01 and HLA-B*57:01 genes that are highly similar, the method differentiated the two genes in all samples. Both purified and unpurified PCR products gave comparable results. This method would be potentially useful as a rapid screening test without the need for purification and denaturation of the PCR products. PMID:27054813

  12. 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

  13. 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.

  14. Multi-step surface functionalization of polyimide based evanescent wave photonic biosensors and application for DNA hybridization by Mach-Zehnder interferometer.

    PubMed

    Melnik, Eva; Bruck, Roman; Hainberger, Rainer; Lämmerhofer, Michael

    2011-08-12

    The process of surface functionalization involving silanization, biotinylation and streptavidin bonding as platform for biospecific ligand immobilization was optimized for thin film polyimide spin-coated silicon wafers, of which the polyimide film serves as a wave guiding layer in evanescent wave photonic biosensors. This type of optical sensors make great demands on the materials involved as well as on the layer properties, such as the optical quality, the layer thickness and the surface roughness. In this work we realized the binding of a 3-mercaptopropyl trimethoxysilane on an oxygen plasma activated polyimide surface followed by subsequent derivatization of the reactive thiol groups with maleimide-PEG(2)-biotin and immobilization of streptavidin. The progress of the functionalization was monitored by using different fluorescence labels for optimization of the chemical derivatization steps. Further, X-ray photoelectron spectroscopy and atomic force microscopy were utilized for the characterization of the modified surface. These established analytical methods allowed to derive information like chemical composition of the surface, surface coverage with immobilized streptavidin, as well as parameters of the surface roughness. The proposed functionalization protocol furnished a surface density of 144 fmol mm(-2) streptavidin with good reproducibility (13.9% RSD, n=10) and without inflicted damage to the surface. This surface modification was applied to polyimide based Mach-Zehnder interferometer sensors to realize a real-time measurement of streptavidin binding validating the functionality of the MZI biosensor. Subsequently, this streptavidin surface was employed to immobilize biotinylated single-stranded DNA and utilized for monitoring of selective DNA hybridization. These proved the usability of polyimide based evanescent photonic devices for biosensing application. PMID:21704776

  15. Quantitative characterization of conformational-specific protein-DNA binding using a dual-spectral interferometric imaging biosensor

    NASA Astrophysics Data System (ADS)

    Zhang, Xirui; Daaboul, George G.; Spuhler, Philipp S.; Dröge, Peter; Ünlü, M. Selim

    2016-03-01

    DNA-binding proteins play crucial roles in the maintenance and functions of the genome and yet, their specific binding mechanisms are not fully understood. Recently, it was discovered that DNA-binding proteins recognize specific binding sites to carry out their functions through an indirect readout mechanism by recognizing and capturing DNA conformational flexibility and deformation. High-throughput DNA microarray-based methods that provide large-scale protein-DNA binding information have shown effective and comprehensive analysis of protein-DNA binding affinities, but do not provide information of DNA conformational changes in specific protein-DNA complexes. Building on the high-throughput capability of DNA microarrays, we demonstrate a quantitative approach that simultaneously measures the amount of protein binding to DNA and nanometer-scale DNA conformational change induced by protein binding in a microarray format. Both measurements rely on spectral interferometry on a layered substrate using a single optical instrument in two distinct modalities. In the first modality, we quantitate the amount of binding of protein to surface-immobilized DNA in each DNA spot using a label-free spectral reflectivity technique that accurately measures the surface densities of protein and DNA accumulated on the substrate. In the second modality, for each DNA spot, we simultaneously measure DNA conformational change using a fluorescence vertical sectioning technique that determines average axial height of fluorophores tagged to specific nucleotides of the surface-immobilized DNA. The approach presented in this paper, when combined with current high-throughput DNA microarray-based technologies, has the potential to serve as a rapid and simple method for quantitative and large-scale characterization of conformational specific protein-DNA interactions.DNA-binding proteins play crucial roles in the maintenance and functions of the genome and yet, their specific binding mechanisms are

  16. Rapid on-site/in-situ detection of heavy metal ions in environmental water using a structure-switching DNA optical biosensor.

    PubMed

    Long, Feng; Zhu, Anna; Shi, Hanchang; Wang, Hongchen; Liu, Jingquan

    2013-01-01

    A structure-switching DNA optical biosensor for rapid on-site/in situ detection of heavy metal ions is reported. Mercury ions (Hg²⁺), highly toxic and ubiquitous pollutants, were selected as model target. In this system, fluorescence-labeled DNA containing T-T mismatch structure was introduced to bind with DNA probes immobilized onto the sensor surface. In the presence of Hg²⁺, some of the fluorescence-labeled DNAs bind with Hg²⁺ to form T-Hg²⁺-T complexes through the folding of themselves into a hairpin structure and dehybridization from the sensor surface, which leads to decrease in fluorescence signal. The total analysis time for a single sample was less than 10 min with detection limit of 1.2 nM. The rapid on-site/in situ determination of Hg²⁺ was readily performed in natural water. This sensing strategy can be extended in principle to other metal ions by substituting the T-Hg²⁺-T complexes with other specificity structures that selectively bind to other analytes. PMID:23892693

  17. Rapid on-site/in-situ detection of heavy metal ions in environmental water using a structure-switching DNA optical biosensor

    NASA Astrophysics Data System (ADS)

    Long, Feng; Zhu, Anna; Shi, Hanchang; Wang, Hongchen; Liu, Jingquan

    2013-07-01

    A structure-switching DNA optical biosensor for rapid on-site/in situ detection of heavy metal ions is reported. Mercury ions (Hg2+), highly toxic and ubiquitous pollutants, were selected as model target. In this system, fluorescence-labeled DNA containing T-T mismatch structure was introduced to bind with DNA probes immobilized onto the sensor surface. In the presence of Hg2+, some of the fluorescence-labeled DNAs bind with Hg2+ to form T-Hg2+-T complexes through the folding of themselves into a hairpin structure and dehybridization from the sensor surface, which leads to decrease in fluorescence signal. The total analysis time for a single sample was less than 10 min with detection limit of 1.2 nM. The rapid on-site/in situ determination of Hg2+ was readily performed in natural water. This sensing strategy can be extended in principle to other metal ions by substituting the T-Hg2+-T complexes with other specificity structures that selectively bind to other analytes.

  18. DNA-Aptamer optical biosensors based on a LPG-SPR optical fiber platform for point-of-care diagnostic

    NASA Astrophysics Data System (ADS)

    Coelho, L.; Queirós, R. B.; Santos, J. L.; Martins, M. Cristina L.; Viegas, D.; Jorge, P. A. S.

    2014-03-01

    Surface Plasmon Resonance (SPR) is the base for some of the most sensitive label free optical fiber biosensors. However, most solutions presented to date require the use of fragile fiber optic structure such as adiabatic tapers or side polished fibers. On the other hand, long-period fiber gratings (LPG) present themselves as an interesting solution to attain an evanescent wave refractive index sensor platform while preserving the optical fiber integrity. The combination of these two approaches constitute a powerful platform that can potentially reach the highest sensitivities as it was recently demonstrated by detailed theoretical study [1, 2]. In this work, a LPG-SPR platform is explored in different configurations (metal coating between two LPG - symmetric and asymmetric) operating in the telecom band (around 1550 nm). For this purpose LPGs with period of 396 μm are combined with tailor made metallic thin films. In particular, the sensing regions were coated with 2 nm of chromium to improve the adhesion to the fiber and 16 nm of gold followed by a 100 nm thick layer of TiO2 dielectric material strategically chosen to attain plasmon resonance in the desired wavelength range. The obtained refractometric platforms were then validated as a biosensor. For this purpose the detection of thrombin using an aptamer based probe was used as a model system for protein detection. The surface of the sensing fibers were cleaned with isopropanol and dried with N2 and then the aminated thrombin aptamer (5'-[NH2]- GGTTGGTGTGGTTGG-3') was immobilized by physisorption using Poly-L-Lysine (PLL) as cationic polymer. Preliminary results indicate the viability of the LPFG-SPR-APTAMER as a flexible platforms point of care diagnostic biosensors.

  19. Accurate and easy-to-use assessment of contiguous DNA methylation sites based on proportion competitive quantitative-PCR and lateral flow nucleic acid biosensor.

    PubMed

    Xu, Wentao; Cheng, Nan; Huang, Kunlun; Lin, Yuehe; Wang, Chenguang; Xu, Yuancong; Zhu, Longjiao; Du, Dan; Luo, Yunbo

    2016-06-15

    Many types of diagnostic technologies have been reported for DNA methylation, but they require a standard curve for quantification or only show moderate accuracy. Moreover, most technologies have difficulty providing information on the level of methylation at specific contiguous multi-sites, not to mention easy-to-use detection to eliminate labor-intensive procedures. We have addressed these limitations and report here a cascade strategy that combines proportion competitive quantitative PCR (PCQ-PCR) and lateral flow nucleic acid biosensor (LFNAB), resulting in accurate and easy-to-use assessment. The P16 gene with specific multi-methylated sites, a well-studied tumor suppressor gene, was used as the target DNA sequence model. First, PCQ-PCR provided amplification products with an accurate proportion of multi-methylated sites following the principle of proportionality, and double-labeled duplex DNA was synthesized. Then, a LFNAB strategy was further employed for amplified signal detection via immune affinity recognition, and the exact level of site-specific methylation could be determined by the relative intensity of the test line and internal reference line. This combination resulted in all recoveries being greater than 94%, which are pretty satisfactory recoveries in DNA methylation assessment. Moreover, the developed cascades show significantly high usability as a simple, sensitive, and low-cost tool. Therefore, as a universal platform for sensing systems for the detection of contiguous multi-sites of DNA methylation without external standards and expensive instrumentation, this PCQ-PCR-LFNAB cascade method shows great promise for the point-of-care diagnosis of cancer risk and therapeutics. PMID:26914373

  20. Quantitative characterization of conformational-specific protein-DNA binding using a dual-spectral interferometric imaging biosensor

    NASA Astrophysics Data System (ADS)

    Zhang, Xirui; Daaboul, George G.; Spuhler, Philipp S.; Dröge, Peter; Ünlü, M. Selim

    2016-03-01

    DNA-binding proteins play crucial roles in the maintenance and functions of the genome and yet, their specific binding mechanisms are not fully understood. Recently, it was discovered that DNA-binding proteins recognize specific binding sites to carry out their functions through an indirect readout mechanism by recognizing and capturing DNA conformational flexibility and deformation. High-throughput DNA microarray-based methods that provide large-scale protein-DNA binding information have shown effective and comprehensive analysis of protein-DNA binding affinities, but do not provide information of DNA conformational changes in specific protein-DNA complexes. Building on the high-throughput capability of DNA microarrays, we demonstrate a quantitative approach that simultaneously measures the amount of protein binding to DNA and nanometer-scale DNA conformational change induced by protein binding in a microarray format. Both measurements rely on spectral interferometry on a layered substrate using a single optical instrument in two distinct modalities. In the first modality, we quantitate the amount of binding of protein to surface-immobilized DNA in each DNA spot using a label-free spectral reflectivity technique that accurately measures the surface densities of protein and DNA accumulated on the substrate. In the second modality, for each DNA spot, we simultaneously measure DNA conformational change using a fluorescence vertical sectioning technique that determines average axial height of fluorophores tagged to specific nucleotides of the surface-immobilized DNA. The approach presented in this paper, when combined with current high-throughput DNA microarray-based technologies, has the potential to serve as a rapid and simple method for quantitative and large-scale characterization of conformational specific protein-DNA interactions.DNA-binding proteins play crucial roles in the maintenance and functions of the genome and yet, their specific binding mechanisms are

  1. 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

  2. Multiple signal amplification electrogenerated chemiluminescence biosensors for sensitive protein kinase activity analysis and inhibition.

    PubMed

    Wang, Zonghua; Yan, Zhiyong; Sun, Na; Liu, Yang

    2015-06-15

    A novel electrogenerated chemiluminescence (ECL) biosensor was built for the detection of kinase activity based on multiple signal amplification nanoprobes. In this strategy, the Xanthine oxidase (XOD) and 5'-phosphate group end DNA conjugated AuNPs was integrated with the phosphorylated peptide by Zr(4+). The XOD on gold nanoparticles can catalyze dissolved oxygen to produce H2O2 in the presence of hypoxanthine (HA) which acts as a coreactor for luminol ECL reaction. In addition, due to the excellent catalytic activity of gold nanoparticle toward luminol ECL reaction and its large surface area that can accommodate large number of XOD and DNA on the surface, the ECL signal of luminol was significantly amplified, affording a highly sensitive ECL analysis of kinase activity. The as-proposed biosensor presents a low detection limit of 0.09 U mL(-1) for protein kinase A (PKA) activity, wide linear range (from 0.1 to 10 U mL(-1)) and excellent stability even in serum samples. This biosensor can also be applied for quantitative kinase inhibitor evaluation. The robust ECL biosensor provides a valuable tool for the high throughput assay in the applications of clinic diagnostic and therapeutic. PMID:25682506

  3. Piezotronic Effect Enhanced Label-Free Detection of DNA Using a Schottky-Contacted ZnO Nanowire Biosensor.

    PubMed

    Cao, Xiaotao; Cao, Xia; Guo, Huijuan; Li, Tao; Jie, Yang; Wang, Ning; Wang, Zhong Lin

    2016-08-23

    A sensitive and in situ selective label-free DNA sensor based on a Schottky-contacted ZnO nanowire (NW) device has been developed and utilized to detect the human immunodeficiency virus 1 gene in this work. Piezotronic effect on the performance of the DNA sensor is studied by measuring its output current under different compressive strains and target complementary DNA concentrations. By applying a -0.59% compressive strain to a ZnO NW-based DNA sensor, the relative current response is greatly enhanced by 454%. A theoretical model is proposed to explain the observed behaviors of the DNA sensor. This study provides a piezotronically modified method to effectively improve the overall performance of the Schottky-contacted ZnO NW-based DNA sensor. PMID:27478905

  4. DNA biosensor-based on fluorescence detection of E. coli O157:H7 by Au@Ag nanorods.

    PubMed

    Sun, Jiadi; Ji, Jian; Sun, Yanqing; Abdalhai, Mandour H; Zhang, Yinzhi; Sun, Xiulan

    2015-08-15

    A novel DNA sensor for the detection of the Escherichia coli O157:H7 (E. coli O157:H7) eaeA gene was constructed using surface enhanced fluorescence (SEF). The spacing distance dependence nature of Au@Ag nanorods surface enhanced fluorescence was investigated when the cy3-labled single strand DNA(ssDNA) and the stem-loop DNA probe modified on the nanorods was co-hybridized. The result revealed that the fluorescence intensity reached the maximum value with the spacing distance of about 10nm between cy3 and the Au@Ag nanorods surface. Based on this result, a fluorescence "ON/OFF" switch for detecting the eaeA gene of E. coli O157:H7 was constructed. Under optimal conditions, the DNA sensor produced a linear range from 10(-17) to 10(-11) M with a correlation coefficient of 0.9947 and a detection limit of 3.33×10(-18) M, and was also found to be specific in targeting eaeA. The DNA sensor demonstrated a new strategy of combining eaeA recognition and Au@Ag nanorods for fluorescence signal enhancement, and increased sensitivity in the detection of bacterial specific genes. PMID:25829221

  5. Highly sensitive and multiple DNA biosensor based on isothermal strand-displacement polymerase reaction and functionalized magnetic microparticles.

    PubMed

    Luo, Ming; Li, Ningxing; Liu, Yufei; Chen, Chaohui; Xiang, Xia; Ji, Xinghu; He, Zhike

    2014-05-15

    A universal, highly sensitive and selective chemiluminescence (CL) imaging method has been developed for high throughput detection of DNA. After molecular beacon (MB) hybridized with the target DNA, the biotin-labeled primer was attached to a magnetic microparticle (MMP) surface by hybridization with the stem part of the MB to initiate a polymerization of DNA strand, which led to the release of the target and another polymerization cycle. Thus the polymerization produced the multiplication of biotin-labeled primer on the surface of MMPs. Sequentially, the horseradish peroxidase (HRP) was conjugated to MMPs surface through the biotin-streptavidin reaction. Then, the conjugated HRP was determined by the CL imaging method. This proposed method could detect the sequence-specific DNA as low as 0.4 pM and discriminate perfectly matched target DNA from the mismatch DNAs. All in all, this proposed method exhibited an efficient amplification performance, and would open new opportunities for sensitive and high throughput detection of DNA. PMID:24412765

  6. Potential of cross-priming amplification and DNA-based lateral-flow strip biosensor for rapid on-site GMO screening.

    PubMed

    Huang, Xin; Zhai, Congcong; You, Qimin; Chen, Hongjun

    2014-07-01

    The requirement to monitor the presence of genetically modified organisms (GMO) in a variety of marked products has generated an increasing demand for reliable, rapid, and time and cost-effective analytical methods. Here we report an on-site method for rapid detection of cauliflower mosaic virus promoter (CaMV 35S), a common element present in most GMO, using cross-priming amplification (CPA) technology. Detection was achieved using a DNA-based contamination-proof strip biosensor. The limit of detection was 30 copies for the pBI121 plasmid containing the CaMV 35S gene. The certified reference sample of GM maize line MON810 was detectable even at the low relative mass concentration of 0.05%. The developed CPA method had high specificity for the CaMV 35S gene, as compared with other GM lines not containing this gene and non-GM products. The method was further validated using nine real-world samples, and the results were confirmed by real-time PCR analysis. Because of its simplicity, rapidity, and high sensitivity, this method of detecting the CaMV 35S gene has great commercial prospects for rapid GMO screening of high-consumption food and agriculture products. PMID:24736809

  7. Renewable Surface Biosensors with Optical Detection

    SciTech Connect

    Bruckner-Lea, Cindy J.; Ackerman, Eric J.; Dockendorff, Brian P.; Holman, David A.; Grate, Jay W.

    2001-04-30

    One major challenge in the development of biosensors is the limited lifetime of a chemically selective surface that includes biomolecules. Renewable surface biosensors address this issue by using fresh aliquots of derivatized microbeads for each analysis. The analyte detection can then occur on the microbeads, or downstream from the microbeads. In this paper, we will describe two types of renewable surface biosensors. The first renewable biosensor system includes on-column optical detection for monitoring the binding of biomolecules onto protein or DNA-derivatized Sepharose beads. The second renewable biosensor system includes detection downstream from the microparticles and is based on the use of derivatized magnetic particles for selective binding. The magnetic particles are fluidically captured and released in a sequential injection system to allow the automation of an Enzyme Linked ImmunoSorbent Assay.

  8. Renewable Surface Biosensors With Optical Detection

    SciTech Connect

    Bruckner-Lea, Cynthia J.; Ackerman, Eric J.; Dockendorff, Brian P.; Holman, David A.; Grate, Jay W.

    2001-12-01

    One major challenge in the development of biosensors is the limited lifetime of a chemically selective surface that includes biomolecules. Renewable surface biosensors address this issue by using fresh aliquots of derivatized microbeads for each analysis. The analyte detection can then occur on the microbeads, or downstream from the microbeads. In this paper, we will describe two types of renewable surface biosensors. The first renewable biosensor system includes on-column optical detection for monitoring the binding of biomolecules onto protein or DNA-derivatized Sepharose beads. The second renewable biosensor system includes detection downstream from the microparticles and is based on the use of derivatized magnetic particles for selective binding. The magnetic particles are fluidically captured and released in a sequential injection system to allow the automation of an Enzyme Linked ImmunoSorbent Assay.

  9. Label-free DNA biosensor based on a peptide nucleic acid-functionalized microstructured optical fiber-Bragg grating

    NASA Astrophysics Data System (ADS)

    Candiani, Alessandro; Bertucci, Alessandro; Giannetti, Sara; Konstantaki, Maria; Manicardi, Alex; Pissadakis, Stavros; Cucinotta, Annamaria; Corradini, Roberto; Selleri, Stefano

    2013-05-01

    We describe a novel sensing approach based on a functionalized microstructured optical fiber-Bragg grating for specific DNA target sequences detection. The inner surface of a microstructured fiber, where a Bragg grating was previously inscribed, has been functionalized by covalent linking of a peptide nucleic acid probe targeting a DNA sequence bearing a single point mutation implicated in cystic fibrosis (CF) disease. A solution of an oligonucleotide (ON) corresponding to a tract of the CF gene containing the mutated DNA has been infiltrated inside the fiber capillaries and allowed to hybridize to the fiber surface according to the Watson-Crick pairing. In order to achieve signal amplification, ON-functionalized gold nanoparticles were then infiltrated and used in a sandwich-like assay. Experimental measurements show a clear shift of the reflected high order mode of a Bragg grating for a 100 nM DNA solution, and fluorescence measurements have confirmed the successful hybridization. Several experiments have been carried out on the same fiber using the identical concentration, showing the same modulation trend, suggesting the possibility of the reuse of the sensor. Measurements have also been made using a 100 nM mismatched DNA solution, containing a single nucleotide mutation and corresponding to the wild-type gene, and the results demonstrate the high selectivity of the sensor.

  10. "Signal-on" photoelectrochemical biosensor for sensitive detection of human T-Cell lymphotropic virus type II DNA: dual signal amplification strategy integrating enzymatic amplification with terminal deoxynucleotidyl transferase-mediated extension.

    PubMed

    Shen, Qingming; Han, Li; Fan, Gaochao; Zhang, Jian-Rong; Jiang, Liping; Zhu, Jun-Jie

    2015-01-01

    A novel "signal-on" photoelectrochemical (PEC) biosensor for sensitive detection of human T-cell lymphotropic virus type II (HTLV-II) DNA was developed on the basis of enzymatic amplification coupled with terminal deoxynucleotidyl transferase (TdT)-mediated extension strategy. The intensity of the photocurrent signal was proportional to the concentration of the HTLV-II DNA-target DNA (tDNA) by dual signal amplification. In this protocol, GR-CdS:Mn/ZnS nanocomposites were used as photoelectric conversion material, while pDNA was used as the tDNA recognizing unit. Moreover, the TdT-mediated extension and the enzymatic signal amplification technique were used to enhance the sensitivity of detection. Using this novel dual signal amplification strategy, the prototype of PEC DNA sensor can detect as low as ∼0.033 fM of HTLV-II DNA with a linear range of 0.1-5000 fM, with excellent differentiation ability even for single-base mismatches. This PEC DNA assay opens a promising platform to detect various DNA targets at ultralow levels for early diagnoses of different diseases. PMID:25871300

  11. 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

  12. Indole-3-acetic acid biosensor based on G-rich DNA labeled AuNPs as chemiluminescence probe coupling the DNA signal amplification

    NASA Astrophysics Data System (ADS)

    Hun, Xu; Mei, Zhenghua; Wang, Zhouping; He, Yunhua

    2012-09-01

    A highly sensitive chemiluminescence (CL) method for detection of phytohormone indole-3-acetic acid (IAA) was developed by using G-rich DNA labeled gold nanoparticles (AuNPs) as CL probe coupling the DNA signal amplification technology. The IAA antibody was immobilized on carboxyl terminated magnetic beads (MBs). In the presence of IAA, antibody labeled AuNPs were captured by antibody functionalized MBs. The DNA on AuNPs is released by a ligand exchange process induced by the addition of DTT. The released DNA is then acted as the linker and hybridized with the capture DNA on MBs and probe DNA on AuNPs CL probe. The CL signal is obtained via the instantaneous derivatization reaction between a specific CL reagent, 3,4,5-trimethoxyl-phenylglyoxal (TMPG), and the G-rich DNA on AuNPs CL probe. IAA can be detected in the concentration range from 0.02 ng/mL to 30 ng/mL, and the limit of detection is 0.01 ng/mL.

  13. 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.

  14. A novel fluorescent biosensor for detection of target DNA fragment from the transgene cauliflower mosaic virus 35S promoter.

    PubMed

    Qiu, Bin; Zhang, Ya-shan; Lin, Yi-bing; Lu, Yu-Jing; Lin, Zhen-yu; Wong, Kwok-Yin; Chen, Guo-nan

    2013-03-15

    In this paper, we reported a convenient fluorescence method for the detection of genetically modified organisms (GMOs). As it is known that the cauliflower mosaic virus (CaMV) 35S promoter is widely used in most transgenic plants (Schnurr and Guerra, 2000), we thus design a simple method based on the detection of a section target DNA (DNA-T) from the transgene CaMV 35S promoter. In this method, the full-length guanine-rich single-strand sequences were split into fragments (Probe 1 and 2) and each part of the fragment possesses two GGG repeats. In the presence of K(+) ion and berberine, if a complementary target DNA of the CaMV 35S promoter was introduced to hybridize with Probe 1 and 2, a G-quadruplex-berberine complex was thus formed and generated a strong fluorescence signal. The generation of fluorescence signal indicates the presence of CaMV 35S promoter. This method is able to identify and quantify Genetically Modified Organisms (GMOs), and it shows wide linear ranges from 5.0×10(-9) to 9.0×10(-7) mol/L with a detection limit of 2.0×10(-9) mol/L. PMID:22959013

  15. Fiber optic-based biosensor

    NASA Technical Reports Server (NTRS)

    Ligler, Frances S.

    1991-01-01

    The NRL fiber optic biosensor is a device which measures the formation of a fluorescent complex at the surface of an optical fiber. Antibodies and DNA binding proteins provide the mechanism for recognizing an analyze and immobilizing a fluorescent complex on the fiber surface. The fiber optic biosensor is fast, sensitive, and permits analysis of hazardous materials remote from the instrumentation. The fiber optic biosensor is described in terms of the device configuration, chemistry for protein immobilization, and assay development. A lab version is being used for assay development and performance characterization while a portable device is under development. Antibodies coated on the fiber are stable for up to two years of storage prior to use. The fiber optic biosensor was used to measure concentration of toxins in the parts per billion (ng/ml) range in under a minute. Immunoassays for small molecules and whole bacteria are under development. Assays using DNA probes as the detection element can also be used with the fiber optic sensor, which is currently being developed to detect biological warfare agents, explosives, pathogens, and toxic materials which pollute the environment.

  16. A Metal-Organic Framework/DNA Hybrid System as a Novel Fluorescent Biosensor for Mercury(II) Ion Detection.

    PubMed

    Wu, Lan-Lan; Wang, Zhuo; Zhao, Shu-Na; Meng, Xing; Song, Xue-Zhi; Feng, Jing; Song, Shu-Yan; Zhang, Hong-Jie

    2016-01-11

    Mercury(II) ions have emerged as a widespread environmental hazard in recent decades. Despite different kinds of detection methods reported to sense Hg(2+) , it still remains a challenging task to develop new sensing molecules to replenish the fluorescence-based apparatus for Hg(2+) detection. This communication demonstrates a novel fluorescent sensor using UiO-66-NH2 and a T-rich FAM-labeled ssDNA as a hybrid system to detect Hg(2+) sensitively and selectively. To the best of our knowledge, it has rarely been reported that a MOF is utilized as the biosensing platform for Hg(2+) assay. PMID:26555340

  17. A label-free fluorescent biosensor for ultratrace detection of terbium (ш) based on structural conversion of G-quadruplex DNA mediated by ThT and terbium (ш).

    PubMed

    Chen, Qiang; Zuo, Junfeng; Chen, Jinfeng; Tong, Ping; Mo, Xiujuan; Zhang, Lan; Li, Jianrong

    2015-10-15

    In this paper, a novel label-free fluorescent biosensor for terbium (ш) (Tb(3+)) was proposed based on structural conversion of G-quadruplex DNA mediated by Thioflavin T (ThT) and Tb(3+). In the presence of K(+), ThT could bind to K(+)-stabilized parallel G-quadruplex, giving rise to high fluorescence intensity. Upon the addition of Tb(3+), Tb(3+) could competitively bind to parallel G-quadruplex leading to the structural change, which resulted in fluorescence decrease. The change of fluorescence intensity (ΔF=F0-F) showed a good linear response toward the concentration of Tb(3+) over the range from 1.0 pM to 10.0 µM with a limit of detection of 0.55 pM. This proposed biosensor was simple and cost-effective in design and in operation with ultrahigh sensitivity and selectivity. Thus, the proposed biosensor could be a promising candidate for monitoring ultratrace Tb(3+) in environment. PMID:26002017

  18. 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

  19. A novel DNA biosensor integrated with Polypyrrole/streptavidin and Au-PAMAM-CP bionanocomposite probes to detect the rs4839469 locus of the vangl1 gene for dysontogenesis prediction.

    PubMed

    Li, Qingying; Yu, Chao; Gao, Rufei; Xia, Chunyong; Yuan, Guolin; Li, Yuliang; Zhao, Yilin; Chen, Qiutong; He, Junlin

    2016-06-15

    The single nucleotide polymorphism (SNP) of the vangl1 gene is highly correlated with Neural Tube Defects (NTDs), a group of severe congenital malformations. It is hindered by the lack of a quantitative detection method. We first propose the use of a DNA biosensor to detect the missense single nucleotide polymorphism (rs4839469 c.346G>A p.Ala116Thr) of the vangl1 gene in this work. Polypyrrole (PPy) and streptavidin were integrated to modify a gold electrode. We took advantage of the PPy's good biocompatibility and excellent conductivity. To further accelerate the electron transfer process at the electrode surface, polyamidoamine dendrimer-encapsulated gold nanoparticles (Au-PAMAM) were used, because Au-PAMAM possess a large number of amino groups to load capture probes (CP). Using the biotin-streptavidin system, the Au-PAMAM-CP bionanocomposite probe, which can detect the target DNA, was conjugated to the electrode surface. Under optimal conditions, the DNA biosensor exhibited a wide linear range of 0.1-100nM with a low detection limit of 0.033nM (S/N=3). The results suggest that this approach has the potential to be used in clinical research. PMID:26914375

  20. 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.

  1. Label-Free Impedance Biosensors: Opportunities and Challenges

    PubMed Central

    Daniels, Jonathan S.; Pourmand, Nader

    2007-01-01

    Impedance biosensors are a class of electrical biosensors that show promise for point-of-care and other applications due to low cost, ease of miniaturization, and label-free operation. Unlabeled DNA and protein targets can be detected by monitoring changes in surface impedance when a target molecule binds to an immobilized probe. The affinity capture step leads to challenges shared by all label-free affinity biosensors; these challenges are discussed along with others unique to impedance readout. Various possible mechanisms for impedance change upon target binding are discussed. We critically summarize accomplishments of past label-free impedance biosensors and identify areas for future research. PMID:18176631

  2. Meditating metal coenhanced fluorescence and SERS around gold nanoaggregates in nanosphere as bifunctional biosensor for multiple DNA targets.

    PubMed

    Liu, Yong; Wu, Peiyi

    2013-06-26

    Gold nanoparticles (Au NPs) are very attractive candidate nanoparticles in biological assay because of their high chemical stabilities, high homogeneities, good biocompatibilities, and low toxicities. However, molecular beacon assays via encapsulating the combined fluorescence or surface-enhanced Raman scattering (SERS) signals of reporters and Au NPs in nanobarcodes particles usually suffer from fluorescence quenching or weak Raman enhancement when Au NPs are employed (especially with size smaller than 15 nm). Herein, we present a new design of simultaneously realizing metal-enhanced fluorescence and coenhanced surface-enhanced Raman scattering by facilely embedding Ag nanoparticle into the shell of two kinds of Au nanoaggregate (5 and 10 nm), meanwhile, fluorophore is located between the silver core and gold nanoparticle layers and the distance among them is adjusted by SiO2 spacer (Ag@first SiO2 spacer@FiTC+SiO2@second SiO2 spacer@Au nanoaggregate). In this architecture, Ag nanoparticle not only is utilized as an efficient fluorescence enhancer to overcome the common fluorescence quenching around Au nanoaggregates but also behaves like a mirror. Thus, incident light that passes through the SERS-active Au nanoaggregate and the intervening dielectric layer of SiO2 could be reflected multiply from the surface of Ag nanoparticle and coupled with the light at the nanogap between the Au nanoaggregates to further amplify Raman intensity. This results in enhancement factors for fluorescence and SERS ~1.6-fold and more than 300-fold higher than the control samples without silver core under identical experimental conditions, respectively. Moreover, fluorophore and SERS reporters are assembled onto different layers of the concentric hybrid microsphere, resulting in a feasible fabrication protocol when a large number of agents need to be involved into the dual-mode nanobarcodes. A proof-of-concept chip-based DNA sandwich hybridization assay using genetically modified

  3. 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

  4. 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

  5. Biosensors in clinical chemistry: An overview.

    PubMed

    Murugaiyan, Sathish Babu; Ramasamy, Ramesh; Gopal, Niranjan; Kuzhandaivelu, V

    2014-01-01

    Biosensors are small devices that employ biological/biochemical reactions for detecting target analytes. Basically, the device consists of a biocatalyst and a transducer. The biocatalyst may be a cell, tissue, enzyme or even an oligonucleotide. The transducers are mainly amperometric, potentiometric or optical. The classification of biosensors is based on (a) the nature of the recognition event or (b) the intimacy between the biocatalyst and the transducer. Bioaffinity and biocatalytic devices are examples for the former and the first, whereas second and third generation instruments are examples for the latter. Cell-based biosensors utilizing immobilized cells, tissues as also enzyme immunosensors and DNA biosensors find variegated uses in diagnostics. Enzyme nanoparticle-based biosensors make use of small particles in the nanometer scale and are currently making a mark in laboratory medicine. Nanotechnology can help in optimizing the diagnostic biochips, which would facilitate sensitive, rapid, accurate and precise bedside monitoring. Biosensors render themselves as capable diagnostic tools as they meet most of the above-mentioned criteria. PMID:24627875

  6. 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

  7. Design and application of genetically encoded biosensors

    PubMed Central

    Palmer, Amy E.; Qin, Yan; Park, Jungwon Genevieve; McCombs, Janet E.

    2012-01-01

    In the past 5–10 years, the power of the green fluorescent protein (GFP) and its numerous derivatives has been harnessed toward the development of genetically encoded fluorescent biosensors. These sensors are incorporated into cells or organisms as plasmid DNA, which leads the transcriptional and translational machinery of the cell to express a functional sensor. To date, over 100 different genetically encoded biosensors have been developed for targets as diverse as ions, molecules and enzymes. Such sensors are instrumental in providing a window into the real-time biochemistry of living cells and whole organisms, and are providing unprecedented insight into the inner workings of a cell. PMID:21251723

  8. Plasmonic Nanostructures for Biosensor Applications

    NASA Astrophysics Data System (ADS)

    Gadde, Akshitha

    Improving the sensitivity of existing biosensors is an active research topic that cuts across several disciplines, including engineering and biology. Optical biosensors are the one of the most diverse class of biosensors which can be broadly categorized into two types based on the detection scheme: label-based and label-free detection. In label-based detection, the target bio-molecules are labeled with dyes or tags that fluoresce upon excitation, indicating the presence of target molecules. Label-based detection is highly-sensitive, capable of single molecule detection depending on the detector type used. One method of improving the sensitivity of label-based fluorescence detection is by enhancement of the emission of the labels by coupling them with metal nanostructures. This approach is referred as plasmon-enhanced fluorescence (PEF). PEF is achieved by increasing the electric field around the nano metal structures through plasmonics. This increased electric field improves the enhancement from the fluorophores which in turn improves the photon emission from the fluorophores which, in turn, improves the limit of detection. Biosensors taking advantage of the plasmonic properties of metal films and nanostructures have emerged an alternative, low-cost, high sensitivity method for detecting labeled DNA. Localized surface plasmon resonance (LSPR) sensors employing noble metal nanostructures have recently attracted considerable attention as a new class of plasmonic nanosensors. In this work, the design, fabrication and characterization of plasmonic nanostructures is carried out. Finite difference time domain (FDTD) simulations were performed using software from Lumerical Inc. to design a novel LSPR structure that exhibit resonance overlapping with the absorption and emission wavelengths of quantum dots (QD). Simulations of a composite Au/SiO2 nanopillars on silicon substrate were performed using FDTD software to show peak plasmonic enhancement at QD emission wavelength

  9. Graphene-Based Optical Biosensors and Imaging

    SciTech Connect

    Tang, Zhiwen; He, Shijiang; Pei, Hao; Du, Dan; Fan, Chunhai; Lin, Yuehe

    2014-01-13

    This chapter focuses on the design, fabrication and application of graphene based optical nanobiosensors. The emerging graphene based optical nanobiosensors demonstrated the promising bioassay and biomedical applications thanking to the unique optical features of graphene. According to the different applications, the graphene can be tailored to form either fluorescent emitter or efficient fluorescence quencher. The exceptional electronic feature of graphene makes it a powerful platform for fabricating the SPR and SERS biosensors. Today the graphene based optical biosensors have been constructed to detect various targets including ions, small biomolecules, DNA/RNA and proteins. This chapter reviews the recent progress in graphene-based optical biosensors and discusses the opportunities and challenges in this field.

  10. Dynamic Modulation of DNA Hybridization Using Allosteric DNA Tetrahedral Nanostructures.

    PubMed

    Song, Ping; Li, Min; Shen, Juwen; Pei, Hao; Chao, Jie; Su, Shao; Aldalbahi, Ali; Wang, Lihua; Shi, Jiye; Song, Shiping; Wang, Lianhui; Fan, Chunhai; Zuo, Xiaolei

    2016-08-16

    The fixed dynamic range of traditional biosensors limits their utility in several real applications. For example, viral load monitoring requires the dynamic range spans several orders of magnitude; whereas, monitoring of drugs requires extremely narrow dynamic range. To overcome this limitation, here, we devised tunable biosensing interface using allosteric DNA tetrahedral bioprobes to tune the dynamic range of DNA biosensors. Our strategy takes the advantage of the readily and flexible structure design and predictable geometric reconfiguration of DNA nanotechnology. We reconfigured the DNA tetrahedral bioprobes by inserting the effector sequence into the DNA tetrahedron, through which, the binding affinity of DNA tetrahedral bioprobes can be tuned. As a result, the detection limit of DNA biosensors can be programmably regulated. The dynamic range of DNA biosensors can be tuned (narrowed or extended) for up to 100-fold. Using the regulation of binding affinity, we realized the capture and release of biomolecules by tuning the binding behavior of DNA tetrahedral bioprobes. PMID:27435955

  11. Biosentinel: Developing a Space Radiation Biosensor

    NASA Technical Reports Server (NTRS)

    Santa Maria, Sergio R.; Marina, Diana B.; Parra, Macarena P.; Boone, Travis D.; Tan, Ming; Ricco, Antonio J.; Straume, Tore N.; Lusby, Terry C.; Harkness, T.; Reiss-Bubenheim, Debra; Brent, R.; Bhattacharya, Sharmila

    2014-01-01

    Ionizing radiation presents a major challenge to human exploration and long-term residence in space. The deep-space radiation spectrum includes highly energetic particles that generate double strand breaks (DSBs), deleterious DNA lesions that are usually repaired without errors via homologous recombination (HR), a conserved pathway in all eukaryotes. While progress identifying and characterizing biological radiation effects using Earth-based facilities has been significant, no terrestrial source duplicates the unique space radiation environment.We are developing a biosensor-based nanosatellite to fly aboard NASAs Space Launch System Exploration Mission 1, expected to launch in 2017 and reach a 1AU (astronomic unit) heliocentric orbit. Our biosensor (called BioSentinel) uses the yeast S. cerevisiae to measure DSBs in response to ambient space radiation. The BioSentinel strain contains engineered genetic defects that prevent growth until and unless a radiation-induced DSB near a reporter gene activates the yeasts HR repair mechanisms. Thus, culture growth and metabolic activity directly indicate a successful DSB-and-repair event. In parallel, HR-defective and wild type strains will provide survival data. Desiccated cells will be carried within independent culture microwells, built into 96-well microfluidic cards. Each microwell set will be activated by media addition at different time points over 18 months, and cell growth will be tracked continuously via optical density. One reserve set will be activated only in the occurrence of a solar particle event. Biological measurements will be compared to data provided by onboard physical dosimeters and to Earth-based experiments.BioSentinel will conduct the first study of biological response to space radiation outside Low Earth Orbit in over 40 years. BioSentinel will thus address strategic knowledge gaps related to the biological effects of space radiation and will provide an adaptable platform to perform human

  12. Replaceable Microfluidic Cartridges for a PCR Biosensor

    NASA Technical Reports Server (NTRS)

    Francis, Kevin; Sullivan, Ron

    2005-01-01

    The figure depicts a replaceable microfluidic cartridge that is a component of a miniature biosensor that detects target deoxyribonucleic acid (DNA) sequences. The biosensor utilizes (1) polymerase chain reactions (PCRs) to multiply the amount of DNA to be detected, (2) fluorogenic polynucleotide probe chemicals for labeling the target DNA sequences, and (3) a high-sensitivity epifluorescence-detection optoelectronic subsystem. Microfluidics is a relatively new field of device development in which one applies techniques for fabricating microelectromechanical systems (MEMS) to miniature systems for containing and/or moving fluids. Typically, microfluidic devices are microfabricated, variously, from silicon or polymers. The development of microfluidic devices for applications that involve PCR and fluorescence-based detection of PCR products poses special challenges

  13. Biosensors for Cell Analysis.

    PubMed

    Zhou, Qing; Son, Kyungjin; Liu, Ying; Revzin, Alexander

    2015-01-01

    Biosensors first appeared several decades ago to address the need for monitoring physiological parameters such as oxygen or glucose in biological fluids such as blood. More recently, a new wave of biosensors has emerged in order to provide more nuanced and granular information about the composition and function of living cells. Such biosensors exist at the confluence of technology and medicine and often strive to connect cell phenotype or function to physiological or pathophysiological processes. Our review aims to describe some of the key technological aspects of biosensors being developed for cell analysis. The technological aspects covered in our review include biorecognition elements used for biosensor construction, methods for integrating cells with biosensors, approaches to single-cell analysis, and the use of nanostructured biosensors for cell analysis. Our hope is that the spectrum of possibilities for cell analysis described in this review may pique the interest of biomedical scientists and engineers and may spur new collaborations in the area of using biosensors for cell analysis. PMID:26274599

  14. Introduction to biosensors

    PubMed Central

    Bhalla, Nikhil; Jolly, Pawan; Formisano, Nello

    2016-01-01

    Biosensors are nowadays ubiquitous in biomedical diagnosis as well as a wide range of other areas such as point-of-care monitoring of treatment and disease progression, environmental monitoring, food control, drug discovery, forensics and biomedical research. A wide range of techniques can be used for the development of biosensors. Their coupling with high-affinity biomolecules allows the sensitive and selective detection of a range of analytes. We give a general introduction to biosensors and biosensing technologies, including a brief historical overview, introducing key developments in the field and illustrating the breadth of biomolecular sensing strategies and the expansion of nanotechnological approaches that are now available. PMID:27365030

  15. Introduction to biosensors.

    PubMed

    Bhalla, Nikhil; Jolly, Pawan; Formisano, Nello; Estrela, Pedro

    2016-06-30

    Biosensors are nowadays ubiquitous in biomedical diagnosis as well as a wide range of other areas such as point-of-care monitoring of treatment and disease progression, environmental monitoring, food control, drug discovery, forensics and biomedical research. A wide range of techniques can be used for the development of biosensors. Their coupling with high-affinity biomolecules allows the sensitive and selective detection of a range of analytes. We give a general introduction to biosensors and biosensing technologies, including a brief historical overview, introducing key developments in the field and illustrating the breadth of biomolecular sensing strategies and the expansion of nanotechnological approaches that are now available. PMID:27365030

  16. Toward Hypoxia-Selective DNA-Alkylating Agents Built by Grafting Nitrogen Mustards onto the Bioreductively Activated, Hypoxia-Selective DNA-Oxidizing Agent 3-Amino-1,2,4-benzotriazine 1,4-Dioxide (Tirapazamine)

    PubMed Central

    2015-01-01

    Tirapazamine (3-amino-1,2,4-benzotriazine 1,4-dioxide) is a heterocyclic di-N-oxide that undergoes enzymatic deoxygenation selectively in the oxygen-poor (hypoxic) cells found in solid tumors to generate a mono-N-oxide metabolite. This work explored the idea that the electronic changes resulting from the metabolic deoxygenation of tirapazamine analogues might be exploited to activate a DNA-alkylating species selectively in hypoxic tissue. Toward this end, tirapazamine analogues bearing nitrogen mustard units were prepared. In the case of the tirapazamine analogue 18a bearing a nitrogen mustard unit at the 6-position, it was found that removal of the 4-oxide from the parent di-N-oxide to generate the mono-N-oxide analogue 17a did indeed cause a substantial increase in reactivity of the mustard unit, as measured by hydrolysis rates and DNA-alkylation yields. Hammett sigma values were measured to quantitatively assess the magnitude of the electronic changes induced by metabolic deoxygenation of the 3-amino-1,2,4-benzotriazine 1,4-dioxide heterocycle. The results provide evidence that the 1,2,4-benzotiazine 1,4-dioxide unit can serve as an oxygen-sensing prodrug platform for the selective unmasking of bioactive agents in hypoxic cells. PMID:25029663

  17. Low-cost and highly efficient DNA biosensor for heavy metal ion using specific DNAzyme-modified microplate and portable glucometer-based detection mode.

    PubMed

    Zhang, Jin; Tang, Ying; Teng, Liumei; Lu, Minghua; Tang, Dianping

    2015-06-15

    A simple and low-cost DNA sensing platform based on Pb(2+)-specific DNAzyme-modified microplate was successfully developed for highly sensitive monitoring of lead ion (Pb(2+), one kind of toxic heavy metal ion) in the environmental samples coupling with a portable personal glucometer (PGM)-based detection mode. The detection cell was first prepared simply by means of immobilizing the DNAzyme on the streptavidin-modified microplate. Gold nanoparticle labeled with single-stranded DNA and invertase (Enz-AuNP-DNA) was utilized as the signal-transduction tag to produce PGM substrate (glucose). Upon addition of lead ion into the microplate, the substrate strand of the immobilized DNAzyme was catalytically cleaved by target Pb(2+), and the newly generated single-strand DNA in the microplate could hybridize again with the single-stranded DNA on the Enz-AuNP-DNA. Accompanying with the Enz-AuNP-DNA, the carried invertase could convert sucrose into glucose. The as-produced glucose could be monitored by using a widely accessible PGM for in situ amplified digital readout. Based on Enz-AuNP-DNA amplification strategy, as low as 1.0 pM Pb(2+) could be detected under the optimal conditions. Moreover, the methodology also showed good reproducibility and high selectivity toward target Pb(2+) against other metal ions because of highly specific Pb(2+)-dependent DNAzyme, and was applicable for monitoring Pb(2+) in the naturally contaminated sewage and spiked drinking water samples. PMID:25576929

  18. BIOSENSORS FOR ENVIRONMENTAL APPLICATIONS

    EPA Science Inventory

    A review, with 19 references, is given on challenges and possible opportunities for the development of biosensors for environmental monitoring applications. The high cost and slow turnaround times typically associated with the measurement of regulated pollutants clearly indicates...

  19. Triggered optical biosensor

    DOEpatents

    Song, Xuedong; Swanson, Basil I.

    2001-10-02

    An optical biosensor is provided for the detection of a multivalent target biomolecule, the biosensor including a substrate having a bilayer membrane thereon, a recognition molecule situated at the surface, the recognition molecule capable of binding with the multivalent target biomolecule, the recognition molecule further characterized as including a fluorescence label thereon and as being movable at the surface and a device for measuring a fluorescence change in response to binding between the recognition molecule and the multivalent target biomolecule.

  20. High-density fiber optic biosensor arrays

    NASA Astrophysics Data System (ADS)

    Epstein, Jason R.; Walt, David R.

    2002-02-01

    Novel approaches are required to coordinate the immense amounts of information derived from diverse genomes. This concept has influenced the expanded role of high-throughput DNA detection and analysis in the biological sciences. A high-density fiber optic DNA biosensor was developed consisting of oligonucleotide-functionalized, 3.1 mm diameter microspheres deposited into the etched wells on the distal face of a 500 micrometers imaging fiber bundle. Imaging fiber bundles containing thousands of optical fibers, each associated with a unique oligonucleotide probe sequence, were the foundation for an optically connected, individually addressable DNA detection platform. Different oligonucleotide-functionalized microspheres were combined in a stock solution, and randomly dispersed into the etched wells. Microsphere positions were registered from optical dyes incorporated onto the microspheres. The distribution process provided an inherent redundancy that increases the signal-to-noise ratio as the square root of the number of sensors examined. The representative amount of each probe-type in the array was dependent on their initial stock solution concentration, and as other sequences of interest arise, new microsphere elements can be added to arrays without altering the existing detection capabilities. The oligonucleotide probe sequences hybridize to fluorescently-labeled, complementary DNA target solutions. Fiber optic DNA microarray research has included DNA-protein interaction profiles, microbial strain differentiation, non-labeled target interrogation with molecular beacons, and single cell-based assays. This biosensor array is proficient in DNA detection linked to specific disease states, single nucleotide polymorphism (SNP's) discrimination, and gene expression analysis. This array platform permits multiple detection formats, provides smaller feature sizes, and enables sensor design flexibility. High-density fiber optic microarray biosensors provide a fast

  1. Built to disappear.

    PubMed

    Bauer, Siegfried; Kaltenbrunner, Martin

    2014-06-24

    Microelectronics dominates the technological and commercial landscape of today's electronics industry; ultrahigh density integrated circuits on rigid silicon provide the computing power for smart appliances that help us organize our daily lives. Integrated circuits function flawlessly for decades, yet we like to replace smart phones and tablet computers every year. Disposable electronics, built to disappear in a controlled fashion after the intended lifespan, may be one of the potential applications of transient single-crystalline silicon nanomembranes, reported by Hwang et al. in this issue of ACS Nano. We briefly outline the development of this latest branch of electronics research, and we present some prospects for future developments. Electronics is steadily evolving, and 20 years from now we may find it perfectly normal for smart appliances to be embedded everywhere, on textiles, on our skin, and even in our body. PMID:24892500

  2. Creation of carbon nanotube based bioSensors through dielectrophoretic assembly

    NASA Astrophysics Data System (ADS)

    Mani, Nilan S.; Kim, Steve; Annam, Kaushik; Bane, Danielle; Subramanyam, Guru

    2015-08-01

    Due to their excellent electrical, optical, and mechanical properties, nanosized single wall carbon nanotubes (SWNTs) have attracted significant attention as a transducing element in nano-bio sensor research. Controlled assembly, device fabrication, and bio-functionalization of the SWNTs are crucial in creating the sensors. In this study, working biosensor platforms were created using dielectrophoretic assembly of single wall carbon nanotubes (SWNTs) as a bridge between two gold electrodes. SWNTs in a commercial SDS surfactant solution were dispensed in the gap between the two gold electrodes, followed by applying an ac voltage across the two electrodes. The dielectrophoresis aligns the CNTs and forms a bridge between the two electrodes. A copious washing and a subsequent annealing of the devices at 200 °C remove the surfactants and create an excellent semiconducting (p-type) bridge between the two electrodes. A liquid gated field effect transistor (LGFET) was built using DI water as the gate dielectric and the SWNT bridge as the channel. Negative gate voltages of the FET increased the drain current and applying a positive gate voltage of +0.5V depleted the channel of charges and turned the device off. The biosensor was verified using both the two terminal and three terminal devices. Genomic salmon DNA dissolved in DI water was applied on the SWNT bridge in both type of devices. In the two terminal device, the conductance of the bridge dropped by 65x after the binding of the DNA. In the LGFET, the transconductance of the device decreased 2X after the binding of the DNA. The binding of the DNA also suppressed hysteresis in the Drain Current vs Gate Voltage characteristics of the LGFET.

  3. Recent research trends of radio-frequency biosensors for biomolecular detection.

    PubMed

    Lee, Hee-Jo; Yook, Jong-Gwan

    2014-11-15

    This article reviews radio-frequency (RF) biosensors based on passive and/or active devices and circuits. In particular, we focus on RF biosensors designed for detection of various biomolecules such as biotin-streptavidin, DNA hybridization, IgG, and glucose. The performance of these biosensors has been enhanced by the introduction of various sensing schemes with diverse nanomaterials (e.g., carbon nanotubes, graphene oxide, magnetic and gold nanoparticles, etc.). In addition, the RF biosensing platforms that can be associated with an RF active system are discussed. Finally, the challenges of RF biosensors are presented and suggestions are made for their future direction and prospects. PMID:24934746

  4. 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

  5. A photoelectrochemical biosensor for o-aminophenol based on assembling of CdSe and DNA on TiO2 film electrode.

    PubMed

    Yan, Kai; Wang, Rui; Zhang, Jingdong

    2014-03-15

    A novel photoelectrochemical (PEC) biosensing platform was constructed by assembling CdSe quantum dots (QDs) and DNA on liquid phase deposited TiO2 (DNA-CdSe/TiO2) film electrode. The transmission electron microscopy (TEM), scanning electron microscopy (SEM) and X-ray diffraction (XRD) analysis indicated that CdSe QDs were homogeneously assembled on TiO2 film. The UV-visible diffuse reflectance spectra (DRS) showed that CdSe and DNA could effectively enhance the absorption of TiO2 film to visible light. The obtained electrode showed a sensitive PEC response to o-aminophenol (OAP) under visible light irradiation. Due to the interaction between DNA and OAP, the response of OAP was improved by DNA immobilized on the sensing film. Under optimized conditions, the photocurrent was linearly proportional to OAP in the concentration range from 4.0 × 10(-7) to 2.7 × 10(-5) mol L(-1), with a detection limit (3S/N) of 8.0 × 10(-8) mol L(-1). The novel strategy could provide a fast and sensitive method for OAP determination. PMID:24161564

  6. Fiber optic choline biosensor

    NASA Astrophysics Data System (ADS)

    Wang, Hong; Cao, Xiaojian; Jia, Ke; Chai, Xueting; Lu, Hua; Lu, Zuhong

    2001-10-01

    A fiber optic fluorescence biosensor for choline is introduced in this paper. Choline is an important neurotransmitter in mammals. Due to the growing needs for on-site clinical monitoring of the choline, much effect has been devoted to develop choline biosensors. Fiber-optic fluorescence biosensors have many advantages, including miniaturization, flexibility, and lack of electrical contact and interference. The choline fiber-optic biosensor we designed implemented a bifurcated fiber to perform fluorescence measurements. The light of the blue LED is coupled into one end of the fiber as excitation and the emission spectrum from sensing film is monitored by fiber-spectrometer (S2000, Ocean Optics) through the other end of the fiber. The sensing end of the fiber is coated with Nafion film dispersed with choline oxidase and oxygen sensitive luminescent Ru(II) complex (Tris(2,2'-bipyridyl)dichlororuthenium(II), hexahydrate). Choline oxidase catalyzes the oxidation of choline to betaine and hydrogen peroxide while consuming oxygen. The fluorescence intensity of oxygen- sensitive Ru(II) are related to the choline concentration. The response of the fiber-optic sensor in choline solution is represented and discussed. The result indicates a low-cost, high-performance, portable choline biosensor.

  7. Future of biosensors: a personal view.

    PubMed

    Scheller, Frieder W; Yarman, Aysu; Bachmann, Till; Hirsch, Thomas; Kubick, Stefan; Renneberg, Reinhard; Schumacher, Soeren; Wollenberger, Ulla; Teller, Carsten; Bier, Frank F

    2014-01-01

    Biosensors representing the technological counterpart of living senses have found routine application in amperometric enzyme electrodes for decentralized blood glucose measurement, interaction analysis by surface plasmon resonance in drug development, and to some extent DNA chips for expression analysis and enzyme polymorphisms. These technologies have already reached a highly advanced level and need minor improvement at most. The dream of the "100-dollar" personal genome may come true in the next few years provided that the technological hurdles of nanopore technology or of polymerase-based single molecule sequencing can be overcome. Tailor-made recognition elements for biosensors including membrane-bound enzymes and receptors will be prepared by cell-free protein synthesis. As alternatives for biological recognition elements, molecularly imprinted polymers (MIPs) have been created. They have the potential to substitute antibodies in biosensors and biochips for the measurement of low-molecular-weight substances, proteins, viruses, and living cells. They are more stable than proteins and can be produced in large amounts by chemical synthesis. Integration of nanomaterials, especially of graphene, could lead to new miniaturized biosensors with high sensitivity and ultrafast response. In the future individual therapy will include genetic profiling of isoenzymes and polymorphic forms of drug-metabolizing enzymes especially of the cytochrome P450 family. For defining the pharmacokinetics including the clearance of a given genotype enzyme electrodes will be a useful tool. For decentralized online patient control or the integration into everyday "consumables" such as drinking water, foods, hygienic articles, clothing, or for control of air conditioners in buildings and cars and swimming pools, a new generation of "autonomous" biosensors will emerge. PMID:24196315

  8. An electrochemiluminescence biosensor for 8-oxo-7,8-dihydro-2'-deoxyguanosine quantification and DNA repair enzyme activity analysis using a novel bifunctional probe.

    PubMed

    Wu, Yiping; Yang, Xiqiang; Zhang, Bintian; Guo, Liang-Hong

    2015-07-15

    A new electrochemiluminescence (ECL) sensor was developed for 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) quantification and Escherichia coli formamidopyrimidine-DNA glycosylase (FPG) activity assay. The sensor employed a novel spermine conjugated ruthenium tris-(bipyridine) derivative (spermine-Ru) which binds specifically with 8-oxodGuo through a one-step reaction and also acts as an ECL signal reporter. In the sensor, an 8-oxodGuo-containing ds-DNA film was first immobilized on a gold electrode by self-assembly. The DNA film was then incubated with spermine-Ru under oxidative condition for 8-oxodGuo labeling. The ECL intensity was found to correlate with the amount of 8-oxodGuo on the surface and the detection limit was estimated to be about 1 lesion in 500 DNA bases. Addition of FPG resulted in some loss of the signal due to the excision of 8-oxodGuo by the enzyme. An inverse relationship between ECL intensity and FPG concentration was observed in a range from 0 to 4.0U/µL, demonstrating that this sensor could be used for FPG activity assay. A number of metal ions were screened by the sensor for their inhibition effect on FPG activity. Among them, Hg(2+) and methyl Hg(II) shown very potent inhibition, with IC50 values of 4.04µM and 4.34nM respectively. The result may suggest that interference on the DNA repair system could be another mechanism for the high toxicity of MeHg. PMID:25747509

  9. Integrating and amplifying signal from riboswitch biosensors.

    PubMed

    Goodson, Michael S; Harbaugh, Svetlana V; Chushak, Yaroslav G; Kelley-Loughnane, Nancy

    2015-01-01

    Biosensors offer a built-in energy supply and inherent sensing machinery that when exploited correctly may surpass traditional sensors. However, biosensor systems have been hindered by a narrow range of ligand detection capabilities, a relatively low signal output, and their inability to integrate multiple signals. Integration of signals could increase the specificity of the sensor and enable detection of a combination of ligands that may indicate environmental or developmental processes when detected together. Amplifying biosensor signal output will increase detector sensitivity and detection range. Riboswitches offer the potential to widen the diversity of ligands that may be detected, and advances in synthetic biology are illuminating myriad possibilities in signal processing using an orthogonal parts-based engineering approach. In this chapter, we describe the design, building, and testing of a riboswitch-based Boolean logic AND gate in bacteria, where an output requires the activation of two riboswitches, and the biological circuitry required to amplify the output of the AND gate using natural extracellular bacterial communication signals to "wire" cells together. PMID:25605381

  10. Recent advances in graphene-based biosensors.

    PubMed

    Kuila, Tapas; Bose, Saswata; Khanra, Partha; Mishra, Ananta Kumar; Kim, Nam Hoon; Lee, Joong Hee

    2011-08-15

    A detailed overview towards the advancement of graphene based biosensors has been reviewed. The large surface area and excellent electrical conductivity of graphene allow it to act as an "electron wire" between the redox centers of an enzyme or protein and an electrode's surface. Rapid electron transfer facilitates accurate and selective detection of biomolecules. This review discusses the application of graphene for the detection of glucose, Cyt-c, NADH, Hb, cholesterol, AA, UA, DA, and H(2)O(2). GO and RGO have been used for the fabrication of heavy metal ion sensors, gas sensors, and DNA sensors. Graphene based FETs have also been discussed in details. In all these cases, the biosensors performed well with low working potentials, high sensitivities, low detection limits, and long-term stabilities. PMID:21683572

  11. Biosensors and other medical and environmental probes

    SciTech Connect

    Jacobson, K.B.

    1996-12-31

    The author presents a overview of work at Oak Ridge National Laboratory directed toward the development of biosensors which can be used to monitor for an array of medical and environmental effects. The article describes the variety of problems which have been addressed by development of such sensors, and the range of staff who have been actively involved in this effort. The first such sensor developed at ORNL was an optical fiber whose end was treated with an antibody which would react with the carcinogen benzo(a)pyrene (BaP). Section titles from the article provide an idea of the breadth of applications addressed: medical telesensors; microcantilevers; detecting cancer and health abnormalities; bioreporters; miniaturized devices; biosensors and DNA analysis; lipids in bacteria and human fingerprints; and anthropometry.

  12. Nuclear reactors built, being built, or planned 1992

    SciTech Connect

    Not Available

    1993-07-01

    Nuclear Reactors Built, Being Built, or Planned contains unclassified information about facilities built, being built, or planned in the United States for domestic use or export as of December 31, 1992. The Office of Scientific and Technical Information, US Department of Energy, gathers this information annually from Washington headquarters and field offices of DOE from the US Nuclear Regulatory Commission (NRC); from the US reactor manufacturers who are the principal nuclear contractors for foreign reactor locations; from US and foreign embassies; and from foreign governmental nuclear departments. Information is presented on five parts: Civilian, Production, Military, Export and Critical Assembly.

  13. 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.

  14. Biosensors for bioprocesses

    SciTech Connect

    Van Brunt, J.

    1987-05-01

    The advent of biosensors has been touted as the marriage of the century - a marriage of microelectronics and biotechnology. But exactly what is a biosensor. Actually, the term is used interchangeably for two sometimes very different classes of devices - those that measure biological molecules and particles and those that use biomolecules as part of the sensing mechanism. The basic conceptual design of a biosensor is simple: a biological receptor is coupled to an electronic tranducer in such a way that the transducer converts biochemical activity at one end into electrical activity at the other. The biological component is usually an enzyme (for selective chemical catalysis) or an antibody (for highly selective binding), although cell membrane receptors, tissue slices, and microbial cells are used as well. The electronic component measures voltage (potentiometric), current (amperometric), light, sound, temperaure, or mass (piezoelectric). Biosensors display several unique features that make them especially attractive. They are small. They are simple to use many procedures require one step, no additional reagents, and no radioactivity. They are portable. And they are inexpensive and perfect for data processing.

  15. Bacterium-Based NO2− Biosensor for Environmental Applications

    PubMed Central

    Nielsen, Michael; Larsen, Lars Hauer; Jetten, Mike S. M.; Revsbech, Niels Peter

    2004-01-01

    A sensitive NO2− biosensor that is based on bacterial reduction of NO2− to N2O and subsequent detection of the N2O by a built-in electrochemical N2O sensor was developed. Four different denitrifying organisms lacking NO3− reductase activity were assessed for use in the biosensor. The relevant physiological aspects examined included denitrifying characteristics, growth rate, NO2− tolerance, and temperature and salinity effects on the growth rate. Two organisms were successfully used in the biosensor. The preferred organism was Stenotrophomonas nitritireducens, which is an organism with a denitrifying pathway deficient in both NO3− and N2O reductases. Alternatively Alcaligenes faecalis could be used when acetylene was added to inhibit its N2O reductase. The macroscale biosensors constructed exhibited a linear NO2− response at concentrations up to 1 to 2 mM. The detection limit was around 1 μM NO2−, and the 90% response time was 0.5 to 3 min. The sensor signal was specific for NO2−, and interference was observed only with NH2OH, NO, N2O, and H2S. The sensor signal was affected by changes in temperature and salinity, and calibration had to be performed in a system with a temperature and an ionic strength comparable to those of the medium analyzed. A broad range of water bodies could be analyzed with the biosensor, including freshwater systems, marine systems, and oxic-anoxic wastewaters. The NO2− biosensor was successfully used for long-term online monitoring in wastewater. Microscale versions of the NO2− biosensor were constructed and used to measure NO2− profiles in marine sediment. PMID:15528518

  16. Nuclear reactors built, being built, or planned 1993

    SciTech Connect

    Not Available

    1993-08-01

    Nuclear Reactors Built, Being Built, or Planned contains unclassified information about facilities built, being built, or planned in the United States for domestic use or export as of December 31, 1993. The Office of Scientific and Technical Information, US Department of Energy, gathers this information annually from Washington headquarters and field offices of DOE; from the US Nuclear Regulatory Commission (NRC); from the US reactor manufacturers who are the principal nuclear embassies; and from foreign governmental nuclear departments. The book consists of three divisions, as follows: (1) a commercial reactor locator map and tables of the characteristic and statistical data that follow; a table of abbreviations; (2) tables of data for reactors operating, being built, or planned; and (3) tables of data for reactors that have been shut down permanently or dismantled. The reactors are subdivided into the following parts: civilian, production, military, export, and critical assembly.

  17. Biosensors and bioelectronics on smartphone for portable biochemical detection.

    PubMed

    Zhang, Diming; Liu, Qingjun

    2016-01-15

    Smartphone has been widely integrated with sensors, such as test strips, sensor chips, and hand-held detectors, for biochemical detections due to its portability and ubiquitous availability. Utilizing built-in function modules, smartphone is often employed as controller, analyzer, and displayer for rapid, real-time, and point-of-care monitoring, which can significantly simplify design and reduce cost of the detecting systems. This paper presents a review of biosensors and bioelectronics on smartphone for portable biochemical detections. The biosensors and bioelectronics based on smartphone can mainly be classified into biosensors using optics, surface plasmon resonance, electrochemistry, and near-field communication. The developments of these biosensors and bioelectronics on smartphone are reviewed along with typical biochemical detecting cases. Sensor strategies, detector attachments, and coupling methods are highlighted to show designs of the compact, lightweight, and low-cost sensor systems. The performances and advantages of these designs are introduced with their applications in healthcare diagnosis, environment monitoring, and food evaluation. With advances in micro-manufacture, sensor technology, and miniaturized electronics, biosensor and bioelectronic devices on smartphone can be used to perform biochemical detections as common and convenient as electronic tag readout in foreseeable future. PMID:26319170

  18. BioSentinel: Mission Development of a Radiation Biosensor to Gauge DNA Damage and Repair Beyond Low Earth Orbit on a 6U Nanosatellite

    NASA Technical Reports Server (NTRS)

    Sanchez, Hugo; Lewis, Brian; Hanel, Robert

    2015-01-01

    We are designing and developing a 6U (10 x 22 x 34 cm; 14 kg) nanosatellite as a secondary payload to fly aboard NASAs Space Launch System (SLS) Exploration Mission (EM) 1, scheduled for launch in late 2017. For the first time in over forty years, direct experimental data from biological studies beyond low Earth orbit (LEO) will be obtained during BioSentinels 12- to 18-month mission. BioSentinel will measure the damage and repair of DNA in a biological organism and allow us to compare that to information from onboard physical radiation sensors. In order to understand the relative contributions of the space environments two dominant biological perturbations, reduced gravity and ionizing radiation, results from deep space will be directly compared to data obtained in LEO (on ISS) and on Earth. These data points will be available for validation of existing biological radiation damage and repair models, and for extrapolation to humans, to assist in mitigating risks during future long-term exploration missions beyond LEO. The BioSentinel Payload occupies 4U of the spacecraft and will utilize the monocellular eukaryotic organism Saccharomyces cerevisiae (yeast) to report DNA double-strand-break (DSB) events that result from ambient space radiation. DSB repair exhibits striking conservation of repair proteins from yeast to humans. Yeast was selected because of 1) its similarity to cells in higher organisms, 2) the well-established history of strains engineered to measure DSB repair, 3) its spaceflight heritage, and 4) the wealth of available ground and flight reference data. The S. cerevisiae flight strain will include engineered genetic defects to prevent growth and division until a radiation-induced DSB activates the yeasts DNA repair mechanisms. The triggered culture growth and metabolic activity directly indicate a DSB and its successful repair. The yeast will be carried in the dry state within the 1-atm PL container in 18 separate fluidics cards with each card

  19. Platforms Formed from a Three-Dimensional Cu-Based Zwitterionic Metal-Organic Framework and Probe ss-DNA: Selective Fluorescent Biosensors for Human Immunodeficiency Virus 1 ds-DNA and Sudan Virus RNA Sequences.

    PubMed

    Yang, Shui-Ping; Chen, Shao-Rui; Liu, Shu-Wen; Tang, Xiao-Yan; Qin, Liang; Qiu, Gui-Hua; Chen, Jin-Xiang; Chen, Wen-Hua

    2015-12-15

    We herein report a water-stable three-dimensional Cu-based metal-organic framework (MOF) 1 supported by a tritopic quaternized carboxylate and 4,4'-dipyridyl sulfide as an ancillary ligand. This MOF exhibits unique pore shapes with aromatic rings, positively charged pyridinium and unsaturated Cu(II) cation centers, free carboxylates, tessellating H2O, and coordinating SO4(2-) on the pore surface. Compound 1 can interact with two carboxyfluorescein (FAM)-labeled single-stranded DNA sequences (probe ss-DNA, delineated as P-DNA) through electrostatic, π-stacking, and/or hydrogen-bonding interactions to form two P-DNA@1 systems, and thus quench the fluorescence of FAM via a photoinduced electron-transfer process. These P-DNA@1 systems can be used as effective fluorescent sensors for human immunodeficiency virus 1 double-stranded DNA and Sudan virus RNA sequences, respectively, with detection limits of 196 and 73 pM, respectively. PMID:26619043

  20. Field-Friendly Tuberculosis Biosensor

    NASA Astrophysics Data System (ADS)

    Proper, Nathan; Stone, Jeremy; Jevsevar, Kristen L.; Scherman, Michael; McNeil, Michael R.; Krapf, Diego

    2010-03-01

    Tuberculosis is a fading threat in the United States, but in the developing world it is still a major health-care concern. With the rising number of cases and lack of resources, there is a desperate need for an affordable, portable detection system. Here, we demonstrate the feasibility of a field-friendly immunological biosensor that utilizes florescence and specialized surface chemistries. We observe fluorescently labeled antibodies as they bind to a glass slide. Slides are treated with biotinylated polyethylene glycol to inhibit non-specific interactions and facilitate the binding of primary antibodies allowing for a high degree of specificity. Solutions of tuberculosis-specific antigens where mixed with fluorescently labeled secondary antibodies and incubated on the treated surfaces. An array of different concentrations of antigens bound to fluorescent tags is then read in an epifluorescnece microscope. This assay was used in the portable detector to show that higher concentrations of bound labeled antigens produce a greater emission when excited by a HeNe laser. Home-built electronics, off-the-shelf optics, and a Si photodiode (PD) were used. The data collected from multiple concentrations show a measurable photocurrent. Work is now underway to incorporate a avalanche (PD), flow-cell technology, in a portable box.

  1. BioSentinel: Mission Development of a Radiation Biosensor to Gauge DNA Damage and Repair Beyond Low Earth Orbit on a 6U Nanosatellite.

    NASA Technical Reports Server (NTRS)

    Lewis, Brian; Hanel, Robert; Bhattacharya, Sharmila; Ricco, Antonion J.; Agasid, Elwood; Reiss-Bubenheim, Debra; Straume, Tore; Parra, Macerena; Boone, Travis; Santa Maria, Sergio; Tan, Ming; Marina, Diana; Friedericks, Charlie; Schooley, Aaron; Wu, Shang; Sorgenfrei, Matthew; Rademacher, Abe; Lusby, Terry; Kuroda, Vanessa; Pires, Craig; Benton, Josh; Forman, Doug; Klamm, Ben; Martinez, Andres; Wickizer, Brittany; Sanchez, Hugo; Swan, Bobbie Gale; Semones, Edward; Wheeler, Scott; Ott, C. Mark; Castro, Sarah

    2015-01-01

    We are designing and developing a "6U" (10 x 22 x 34 cm; 14 kg) nanosatellite as a secondary payload to fly aboard NASA's Space Launch System (SLS) Exploration Mission (EM) 1, scheduled for launch in late 2017. For the first time in over forty years, direct experimental data from biological studies beyond low Earth orbit (LEO) will be obtained during BioSentinel's 12- to 18- month mission. BioSentinel will measure the damage and repair of DNA in a biological organism and allow us to compare that to information from onboard physical radiation sensors. In order to understand the relative contributions of the space environment's two dominant biological perturbations, reduced gravity and ionizing radiation, results from deep space will be directly compared to data obtained in LEO (on ISS) and on Earth. These data points will be available for validation of existing biological radiation damage and repair models, and for extrapolation to humans, to assist in mitigating risks during future long-term exploration missions beyond LEO. The BioSentinel Payload occupies 4U of the spacecraft and will utilize the monocellular eukaryotic organism Saccharomyces cerevisiae (yeast) to report DNA double-strand-break (DSB) events that result from ambient space radiation. DSB repair exhibits striking conservation of repair proteins from yeast to humans. Yeast was selected because of 1) its similarity to cells in higher organisms, 2) the well-established history of strains engineered to measure DSB repair, 3) its spaceflight heritage, and 4) the wealth of available ground and flight reference data. The S. cerevisiae flight strain will include engineered genetic defects to prevent growth and division until a radiation-induced DSB activates the yeast's DNA repair mechanisms. The triggered culture growth and metabolic activity directly indicate a DSB and its successful repair. The yeast will be carried in the dry state within the 1-atm P/L container in 18 separate fluidics cards with each

  2. 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.

  3. Functionalized nanopipettes: toward label-free, single cell biosensors

    PubMed Central

    Actis, Paolo; Mak, Andy C.

    2010-01-01

    Nanopipette technology has been proven to be a label-free biosensor capable of identifying DNA and proteins. The nanopipette can include specific recognition elements for analyte discrimination based on size, shape, and charge density. The fully electrical read-out and the ease and low-cost fabrication are unique features that give this technology an enormous potential. Unlike other biosensing platforms, nanopipettes can be precisely manipulated with submicron accuracy and used to study single cell dynamics. This review is focused on creative applications of nanopipette technology for biosensing. We highlight the potential of this technology with a particular attention to integration of this biosensor with single cell manipulation platforms. PMID:20730113

  4. 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.

  5. 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

  6. Fiber Optic Surface Plasmon Resonance-Based Biosensor Technique: Fabrication, Advancement, and Application.

    PubMed

    Liang, Gaoling; Luo, Zewei; Liu, Kunping; Wang, Yimin; Dai, Jianxiong; Duan, Yixiang

    2016-05-01

    Fiber optic-based biosensors with surface plasmon resonance (SPR) technology are advanced label-free optical biosensing methods. They have brought tremendous progress in the sensing of various chemical and biological species. This review summarizes four sensing configurations (prism, grating, waveguide, and fiber optic) with two ways, attenuated total reflection (ATR) and diffraction, to excite the surface plasmons. Meanwhile, the designs of different probes (U-bent, tapered, and other probes) are also described. Finally, four major types of biosensors, immunosensor, DNA biosensor, enzyme biosensor, and living cell biosensor, are discussed in detail for their sensing principles and applications. Future prospects of fiber optic-based SPR sensor technology are discussed. PMID:27119268

  7. Carbon nanotube biosensors

    PubMed Central

    Tîlmaciu, Carmen-Mihaela; Morris, May C.

    2015-01-01

    Nanomaterials possess unique features which make them particularly attractive for biosensing applications. In particular, carbon nanotubes (CNTs) can serve as scaffolds for immobilization of biomolecules at their surface, and combine several exceptional physical, chemical, electrical, and optical characteristics properties which make them one of the best suited materials for the transduction of signals associated with the recognition of analytes, metabolites, or disease biomarkers. Here we provide a comprehensive review on these carbon nanostructures, in which we describe their structural and physical properties, functionalization and cellular uptake, biocompatibility, and toxicity issues. We further review historical developments in the field of biosensors, and describe the different types of biosensors which have been developed over time, with specific focus on CNT-conjugates engineered for biosensing applications, and in particular detection of cancer biomarkers. PMID:26579509

  8. Carbon Nanotube Biosensors

    NASA Astrophysics Data System (ADS)

    Tilmaciu, Carmen-Mihaela; Morris, May

    2015-10-01

    Nanomaterials possess unique features which make them particularly attractive for biosensing applications. In particular Carbon Nanotubes (CNTs) can serve as scaffolds for immobilization of biomolecules at their surface, and combine several exceptional physical, chemical, electrical and optical characteristics properties which make them one of the best suited materials for the transduction of signals associated with the recognition of analytes, metabolites or disease biomarkers. Here we provide a comprehensive review on these carbon nanostructures, in which we will describe their structural and physical properties, discuss functionalization and cellular uptake, biocompatibility and toxicity issues. We further review historical developments in the field of biosensors, and describe the different types of biosensors which have been developed over time, with specific focus on CNT-conjugates engineered for biosensing applications, and in particular detection of cancer biomarkers.

  9. Towards optoelectronic urea biosensors.

    PubMed

    Pokrzywnicka, Marta; Koncki, Robert; Tymecki, Łukasz

    2015-03-01

    Integration of immobilized enzymes with light-emitting diodes (LEDs) leads to the development of optoelectronic enzyme-based biosensors. In this work, urease, used as a model enzyme, immobilized in the form of an open-tubular microbioreactor or biosensing membrane that has been integrated with two red LEDs. It forms complete, fiberless, miniaturized, and extremely economic biooptoelectronic devices useful for nonstationary measurements under flow analysis conditions. Both enzyme-based biodevices, operating according to the paired emitter detector diode (PEDD) principle, allow relatively fast, highly sensitive, and well-reproducible urea detection in the millimolar range of concentrations. Potential analytical applications of the developed urea bioPEDDs have been announced. Both presented constructions will be easily adapted for the development of other optoelectronic biosensors exploring various enzyme-based schemes of biodetection. PMID:25619983

  10. Micro-algal biosensors.

    PubMed

    Brayner, Roberta; Couté, Alain; Livage, Jacques; Perrette, Catherine; Sicard, Clémence

    2011-08-01

    Fighting against water pollution requires the ability to detect pollutants for example herbicides or heavy metals. Micro-algae that live in marine and fresh water offer a versatile solution for the construction of novel biosensors. These photosynthetic microorganisms are very sensitive to changes in their environment, enabling the detection of traces of pollutants. Three groups of micro-algae are described in this paper: chlorophyta, cyanobacteria, and diatoms. PMID:21626188

  11. Thermoresponsive amperometric glucose biosensor.

    PubMed

    Pinyou, Piyanut; Ruff, Adrian; Pöller, Sascha; Barwe, Stefan; Nebel, Michaela; Alburquerque, Natalia Guerrero; Wischerhoff, Erik; Laschewsky, André; Schmaderer, Sebastian; Szeponik, Jan; Plumeré, Nicolas; Schuhmann, Wolfgang

    2016-03-01

    The authors report on the fabrication of a thermoresponsive biosensor for the amperometric detection of glucose. Screen printed electrodes with heatable gold working electrodes were modified by a thermoresponsive statistical copolymer [polymer I: poly(ω-ethoxytriethylenglycol methacrylate-co-3-(N,N-dimethyl-N-2-methacryloyloxyethyl ammonio) propanesulfonate-co-ω-butoxydiethylenglycol methacrylate-co-2-(4-benzoyl-phenoxy)ethyl methacrylate)] with a lower critical solution temperature of around 28 °C in aqueous solution via electrochemically induced codeposition with a pH-responsive redox-polymer [polymer II: poly(glycidyl methacrylate-co-allyl methacrylate-co-poly(ethylene glycol)methacrylate-co-butyl acrylate-co-2-(dimethylamino)ethyl methacrylate)-[Os(bpy)2(4-(((2-(2-(2-aminoethoxy)ethoxy)ethyl)amino)methyl)-N,N-dimethylpicolinamide)](2+)] and pyrroloquinoline quinone-soluble glucose dehydrogenase acting as biological recognition element. Polymer II bears covalently bound Os-complexes that act as redox mediators for shuttling electrons between the enzyme and the electrode surface. Polymer I acts as a temperature triggered immobilization matrix. Probing the catalytic current as a function of the working electrode temperature shows that the activity of the biosensor is dramatically reduced above the phase transition temperature of polymer I. Thus, the local modulation of the temperature at the interphase between the electrode and the bioactive layer allows switching the biosensor from an on- to an off-state without heating of the surrounding analyte solution. PMID:26702635

  12. DNA.

    ERIC Educational Resources Information Center

    Felsenfeld, Gary

    1985-01-01

    Structural form, bonding scheme, and chromatin structure of and gene-modification experiments with deoxyribonucleic acid (DNA) are described. Indicates that DNA's double helix is variable and also flexible as it interacts with regulatory and other molecules to transfer hereditary messages. (DH)

  13. Nuclear reactors built, being built, or planned, 1991

    SciTech Connect

    Simpson, B.

    1992-07-01

    This document contains unclassified information about facilities built, being built, or planned in the United States for domestic use or export as of December 31, 1991. The book is divided into three major sections: Section 1 consists of a reactor locator map and reactor tables; Section 2 includes nuclear reactors that are operating, being built, or planned; and Section 3 includes reactors that have been shut down permanently or dismantled. Sections 2 and 3 contain the following classification of reactors: Civilian, Production, Military, Export, and Critical Assembly. Export reactor refers to a reactor for which the principal nuclear contractor is an American company -- working either independently or in cooperation with a foreign company (Part 4, in each section). Critical assembly refers to an assembly of fuel and assembly of fuel and moderator that requires an external source of neutrons to initiate and maintain fission. A critical assembly is used for experimental measurements (Part 5).

  14. Nuclear reactors built, being built, or planned 1996

    SciTech Connect

    1997-08-01

    This publication contains unclassified information about facilities, built, being built, or planned in the United States for domestic use or export as of December 31, 1996. The Office of Scientific and Technical Information, U.S. Department of Energy, gathers this information annually from Washington headquarters, and field offices of DOE; from the U.S. Nuclear Regulatory Commission (NRC); from the U. S. reactor manufacturers who are the principal nuclear contractors for foreign reactor locations; from U.S. and foreign embassies; and from foreign governmental nuclear departments. The book consists of three divisions, as follows: (1) a commercial reactor locator map and tables of the characteristic and statistical data that follow; a table of abbreviations; (2) tables of data for reactors operating, being built, or planned; and (3) tables of data for reactors that have been shut down permanently or dismantled.

  15. Carbon Nanotube Biosensors for Space Molecule Detection and Clinical Molecular Diagnostics

    NASA Technical Reports Server (NTRS)

    Han, Jie

    2001-01-01

    Both space molecule detection and clinical molecule diagnostics need to develop ultra sensitive biosensors for detection of less than attomole molecules such as amino acids for DNA. However all the electrode sensor systems including those fabricated from the existing carbon nanotubes, have a background level of nA (nanoAmp). This has limited DNA or other molecule detection to nA level or molecules whose concentration is, much higher than attomole level. A program has been created by NASA and NCI (National Cancer Institute) to exploit the possibility of carbon nanotube based biosensors to solve this problem for both's interest. In this talk, I will present our effort on the evaluation and novel design of carbon nanotubes as electrode biosensors with strategies to minimize background currents while maximizing signal intensity.The fabrication of nanotube electrode arrays, immobilization of molecular probes on nanotube electrodes and in vitro biosensor testing will also be discussed.

  16. High-sensitive label-free biosensors based on single-walled carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Maehashi, Kenzo; Matsumoto, Kazuhiko

    2006-02-01

    DNA hybridization has sensitively been detected using carbon nanotube field-effect transistors (CNTFETs) in real time. After full-complementary DNA introduction, the source-drain current gradually increased while monitoring in real time. Full-complementary DNA with concentration as low as 1 fmol/L solution could be effectively detected. Our CNTFET-based biochip is a promising candidate for the development of an integrated, high-throughput, multiplexed DNA biosensor for medical, forensic and environmental diagnostics.

  17. Development of solution-gated graphene transistor model for biosensors

    NASA Astrophysics Data System (ADS)

    Karimi, Hediyeh; Yusof, Rubiyah; Rahmani, Rasoul; Hosseinpour, Hoda; Ahmadi, Mohammad T.

    2014-02-01

    The distinctive properties of graphene, characterized by its high carrier mobility and biocompatibility, have stimulated extreme scientific interest as a promising nanomaterial for future nanoelectronic applications. In particular, graphene-based transistors have been developed rapidly and are considered as an option for DNA sensing applications. Recent findings in the field of DNA biosensors have led to a renewed interest in the identification of genetic risk factors associated with complex human diseases for diagnosis of cancers or hereditary diseases. In this paper, an analytical model of graphene-based solution gated field effect transistors (SGFET) is proposed to constitute an important step towards development of DNA biosensors with high sensitivity and selectivity. Inspired by this fact, a novel strategy for a DNA sensor model with capability of single-nucleotide polymorphism detection is proposed and extensively explained. First of all, graphene-based DNA sensor model is optimized using particle swarm optimization algorithm. Based on the sensing mechanism of DNA sensors, detective parameters ( I ds and V gmin) are suggested to facilitate the decision making process. Finally, the behaviour of graphene-based SGFET is predicted in the presence of single-nucleotide polymorphism with an accuracy of more than 98% which guarantees the reliability of the optimized model for any application of the graphene-based DNA sensor. It is expected to achieve the rapid, quick and economical detection of DNA hybridization which could speed up the realization of the next generation of the homecare sensor system.

  18. Development of solution-gated graphene transistor model for biosensors.

    PubMed

    Karimi, Hediyeh; Yusof, Rubiyah; Rahmani, Rasoul; Hosseinpour, Hoda; Ahmadi, Mohammad T

    2014-01-01

    : The distinctive properties of graphene, characterized by its high carrier mobility and biocompatibility, have stimulated extreme scientific interest as a promising nanomaterial for future nanoelectronic applications. In particular, graphene-based transistors have been developed rapidly and are considered as an option for DNA sensing applications. Recent findings in the field of DNA biosensors have led to a renewed interest in the identification of genetic risk factors associated with complex human diseases for diagnosis of cancers or hereditary diseases. In this paper, an analytical model of graphene-based solution gated field effect transistors (SGFET) is proposed to constitute an important step towards development of DNA biosensors with high sensitivity and selectivity. Inspired by this fact, a novel strategy for a DNA sensor model with capability of single-nucleotide polymorphism detection is proposed and extensively explained. First of all, graphene-based DNA sensor model is optimized using particle swarm optimization algorithm. Based on the sensing mechanism of DNA sensors, detective parameters (Ids and Vgmin) are suggested to facilitate the decision making process. Finally, the behaviour of graphene-based SGFET is predicted in the presence of single-nucleotide polymorphism with an accuracy of more than 98% which guarantees the reliability of the optimized model for any application of the graphene-based DNA sensor. It is expected to achieve the rapid, quick and economical detection of DNA hybridization which could speed up the realization of the next generation of the homecare sensor system. PMID:24517158

  19. Development of solution-gated graphene transistor model for biosensors

    PubMed Central

    2014-01-01

    The distinctive properties of graphene, characterized by its high carrier mobility and biocompatibility, have stimulated extreme scientific interest as a promising nanomaterial for future nanoelectronic applications. In particular, graphene-based transistors have been developed rapidly and are considered as an option for DNA sensing applications. Recent findings in the field of DNA biosensors have led to a renewed interest in the identification of genetic risk factors associated with complex human diseases for diagnosis of cancers or hereditary diseases. In this paper, an analytical model of graphene-based solution gated field effect transistors (SGFET) is proposed to constitute an important step towards development of DNA biosensors with high sensitivity and selectivity. Inspired by this fact, a novel strategy for a DNA sensor model with capability of single-nucleotide polymorphism detection is proposed and extensively explained. First of all, graphene-based DNA sensor model is optimized using particle swarm optimization algorithm. Based on the sensing mechanism of DNA sensors, detective parameters (Ids and Vgmin) are suggested to facilitate the decision making process. Finally, the behaviour of graphene-based SGFET is predicted in the presence of single-nucleotide polymorphism with an accuracy of more than 98% which guarantees the reliability of the optimized model for any application of the graphene-based DNA sensor. It is expected to achieve the rapid, quick and economical detection of DNA hybridization which could speed up the realization of the next generation of the homecare sensor system. PMID:24517158

  20. ESF AS-BUILT CONFIGURATION

    SciTech Connect

    NA

    2005-03-17

    The calculations contained in this document were developed by the ''Mining Group of the Design & Engineering Organization'' and are intended solely for the use of the ''Design & Engineering Organization'' in its work regarding the subsurface repository. Yucca Mountain Project personnel from the ''Mining Group'' should be consulted before use of the calculations for purposes other than those stated herein or use by individuals other than authorized personnel in the ''Design & Engineering Organization''. The purpose of this calculation is to provide design inputs that can be used to develop an as-built drawing of the Exploratory Studies Facility (ESF) for the planning and development of the subsurface repository. This document includes subsurface as-built surveys, recommendation to complete as-built surveys, and Management and Operating Contractor (M&O) Subsurface Design Drawings as inputs. This calculation is used to provide data and information for an as-built ESF subsurface drawing and is not used in the development of results or conclusions, therefore all inputs are considered as indirect.

  1. Improved Biosensors for Soils

    NASA Astrophysics Data System (ADS)

    Silberg, J. J.; Masiello, C. A.; Cheng, H. Y.

    2014-12-01

    Microbes drive processes in the Earth system far exceeding their physical scale, affecting crop yields, water quality, the mobilization of toxic materials, and fundamental aspects of soil biogeochemistry. The tools of synthetic biology have the potential to significantly improve our understanding of microbial Earth system processes: for example, synthetic microbes can be be programmed to report on environmental conditions that stimulate greenhouse gas production, metal oxidation, biofilm formation, pollutant degradation, and microbe-plant symbioses. However, these tools are only rarely deployed in the lab. This research gap arises because synthetically programmed microbes typically report on their environment by producing molecules that are detected optically (e.g., fluorescent proteins). Fluorescent reporters are ideal for petri-dish applications and have fundamentally changed how we study human health, but their usefulness is quite limited in soils where detecting fluorescence is challenging. Here we describe the construction of gas-reporting biosensors, which release nonpolar gases that can be detected in the headspace of incubation experiments. These constructs can be used to probe microbial processes within soils in real-time noninvasive lab experiments. These biosensors can be combined with traditional omics-based approaches to reveal processes controlling soil microbial behavior and lead to improved environmental management decisions.

  2. 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

  3. Multiplexed Biosensors for Mycotoxins.

    PubMed

    Maragos, Chris M

    2016-07-01

    Significant progress has been made in the development of biosensors that can be used to detect low-MW toxins produced by fungi (mycotoxins). The number of formats that have been investigated is impressive and is an indication of the importance attached to finding easy-to-use, accurate, and rapid methods for detecting these toxins in commodities and foods. This review explores the details of multiplexed biosensors based on many formats, including multiplexed immunoassays, suspension arrays, membrane-based devices (flow-through and immunochromatographic), and planar microarrays. Each assay format has its own strengths and areas that need improvement. Certain formats, such as multiplexed immunochromatographic devices, are well developed and relatively easy to use, and in some cases, commercial products are being sold. Others, such as the suspension arrays and microarrays, are laboratory-based assays that, although more complicated, are also more amenable to a larger scale of multiplexing. The diversity of such efforts and the multitude of formats under investigation suggest that multiple solutions will be found to satisfy the need for multiplexed toxin detection. PMID:27455928

  4. 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

  5. DNA

    ERIC Educational Resources Information Center

    Stent, Gunther S.

    1970-01-01

    This history for molecular genetics and its explanation of DNA begins with an analysis of the Golden Jubilee essay papers, 1955. The paper ends stating that the higher nervous system is the one major frontier of biological inquiry which still offers some romance of research. (Author/VW)

  6. Nuclear reactors built, being built, or planned, 1994

    SciTech Connect

    1995-07-01

    This document contains unclassified information about facilities built, being built, or planned in the United States for domestic use or export as of December 31, 1994. The Office of Scientific and Technical Information, US Department of Energy, gathers this information annually from Washington headquarters and field offices of DOE; from the US Nuclear Regulatory Commission (NRC); from the US reactor manufacturers who are the principal nuclear contractors for foreign reactor locations; from US and foreign embassies; and from foreign governmental nuclear departments. The book consists of three divisions, as follows: a commercial reactor locator map and tables of the characteristic and statistical data that follow; a table of abbreviations; tables of data for reactors operating, being built, or planned; and tables of data for reactors that have been shut down permanently or dismantled. The reactors are subdivided into the following parts: Civilian, Production, Military, Export, and Critical Assembly. Export reactor refers to a reactor for which the principal nuclear contractor is a US company -- working either independently or in cooperation with a foreign company (Part 4). Critical assembly refers to an assembly of fuel and moderator that requires an external source of neutrons to initiate and maintain fission. A critical assembly is used for experimental measurements (Part 5).

  7. Nuclear reactors built, being built, or planned: 1995

    SciTech Connect

    1996-08-01

    This report contains unclassified information about facilities built, being built, or planned in the US for domestic use or export as of December 31, 1995. The Office of Scientific and Technical Information, US Department of Energy, gathers this information annually from Washington headquarters and field offices of DOE; from the US Nuclear Regulatory Commission (NRC); from the US reactor manufacturers who are the principal nuclear contractors for foreign reactor locations; from US and foreign embassies; and from foreign governmental nuclear departments. The book consists of three divisions, as follows: (1) a commercial reactor locator map and tables of the characteristic and statistical data that follow; a table of abbreviations; (2) tables of data for reactors operating, being built, or planned; and (3) tables of data for reactors that have been shut down permanently or dismantled. The reactors are subdivided into the following parts: Civilian, Production, Military, Export, and Critical Assembly. Export reactor refers to a reactor for which the principal nuclear contractor is a US company--working either independently or in cooperation with a foreign company (Part 4). Critical assembly refers to an assembly of fuel and moderator that requires an external source of neutrons to initiate and maintain fission. A critical assembly is used for experimental measurements (Part 5).

  8. Biosensor commercialization strategy - a theoretical approach.

    PubMed

    Lin, Chin-Tsai; Wang, Su-Man

    2005-01-01

    Biosensors are analytical devices, which use biological interactions to provide either qualitative or quantitative results. They are extensively employed in many fields such as clinical diagnosis and biomedicine, military applications, anti-terrorism, farm, garden and veterinary analysis, process control, fermentation control and analysis, pharmaceutical and drug analysis, food and drink production and analysis, pollution control and monitoring, microbiology, bacterial and viral analysis, mining, and industrial and toxic gases. The biosensor market has significantly increased and will be mushrooming in the next decade. The total biosensor market is estimated to be 10.8 billion dollars by 2007. The emerging biosensor market presents both opportunities and obstacles to start-up biosensor entrepreneurs. The major challenge and threat for these entrepreneurs is how to predict the biosensor market and how to convert promising biosensor technology into commercialized biosensors. By adopting a simple commercialization strategy framework, we identify two key elements of biosensor commercialization strategy: excludability and complementary asset. We further divide biosensor commercialization environments into four distinct sub-environments: the Attacker's Advantage, Reputation-Based Idea Trading, Greenfield Competition and Ideas Factories. This paper explains how the interaction between these two key elements shapes biosensor commercialization strategy and biosensor industry dynamics. This paper also discusses alternative commercialization strategies for each specific commercialization environment and how to choose from these alternatives. The analysis of this study further provides a good reference for start-up biosensor entrepreneurs to formulate effective biosensor commercialization strategy. PMID:15574353

  9. Built Environment Wind Turbine Roadmap

    SciTech Connect

    Smith, J.; Forsyth, T.; Sinclair, K.; Oteri, F.

    2012-11-01

    The market currently encourages BWT deployment before the technology is ready for full-scale commercialization. To address this issue, industry stakeholders convened a Rooftop and Built-Environment Wind Turbine Workshop on August 11 - 12, 2010, at the National Wind Technology Center, located at the U.S. Department of Energy’s National Renewable Energy Laboratory in Boulder, Colorado. This report summarizes the workshop.

  10. 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.

  11. R&D 100 Winner 2010: Acoustic Wave Biosensors

    ScienceCinema

    Larson, Richard; Branch, Darren; Edwards, Thayne

    2016-06-29

    The acoustic wave biosensor is innovative device that is a handheld, battery-powered, portable detection system capable of multiplex identification of a wide range of medically relevant pathogens and their biomolecular signatures ? viruses, bacteria, proteins, and DNA ? at clinically relevant levels. This detection occurs within minutes ? not hours ? at the point of care, whether that care is in a physician's office, a hospital bed, or at the scene of a biodefense or biomedical emergency.

  12. 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

  13. Noninvasive biosensor for hypoglycemia

    NASA Astrophysics Data System (ADS)

    Varadan, Vijay K.; Whitchurch, Ashwin K.; Sarukesi, Karunakaran

    2003-01-01

    Hypoglycemia-abnormal decrease in blood sugar- is a major obstacle in the management of diabetes and prevention of long-term complications, and it may impose serious effects on the brain, including impairment of memory and other cognitive functions. This is especially a concern in early childhood years when the nervous system is still developing. Hypoglycemic unawareness (in which the body"s normal ability to signal low blood sugar doesn"t work and an oncoming low blood sugar episode proceeds undetected) is a particularly frightening problem for many people with diabetes. Researchers have now uncovered evidence that repeated bouts of insulin-induced hypoglycemia can harm the brain over time, causing confusion, abnormal behavior, loss of consciousness, and seizures. Extreme cases have resulted in coma and death. In this paper, a non-invasive biosensor in a wrist watch along with a wireless data downloading system is proposed.

  14. Slotted photonic crystal biosensors

    NASA Astrophysics Data System (ADS)

    Scullion, Mark Gerard

    Optical biosensors are increasingly being considered for lab-on-a-chip applications due to their benefits such as small size, biocompatibility, passive behaviour and lack of the need for fluorescent labels. The light guiding mechanisms used by many of them result in poor overlap of the optical field with the target molecules, reducing the maximum sensitivity achievable. This thesis presents a new platform for optical biosensors, namely slotted photonic crystals, which engender higher sensitivities due to their ability to confine, spatially and temporally, the peak of optical mode within the analyte itself. Loss measurements showed values comparable to standard photonic crystals, confirming their ability to be used in real devices. A novel resonant coupler was designed, simulated, and experimentally tested, and was found to perform better than other solutions within the literature. Combining with cavities, microfluidics and biological functionalization allowed proof-of-principle demonstrations of protein binding to be carried out. High sensitivities were observed in smaller structures than most competing devices in the literature. Initial tests with cellular material for real applications was also performed, and shown to be of promise. In addition, groundwork to make an integrated device that includes the spectrometer function was also carried out showing that slotted photonic crystals themselves can be used for on-chip wavelength specific filtering and spectroscopy, whilst gas-free microvalves for automation were also developed. This body of work presents slotted photonic crystals as a realistic platform for complete on-chip biosensing; addressing key design, performance and application issues, whilst also opening up exciting new ideas for future study.

  15. Rapid construction of metabolite biosensors using domain-insertion profiling

    PubMed Central

    Nadler, Dana C.; Morgan, Stacy-Anne; Flamholz, Avi; Kortright, Kaitlyn E.; Savage, David F.

    2016-01-01

    Single-fluorescent protein biosensors (SFPBs) are an important class of probes that enable the single-cell quantification of analytes in vivo. Despite advantages over other detection technologies, their use has been limited by the inherent challenges of their construction. Specifically, the rational design of green fluorescent protein (GFP) insertion into a ligand-binding domain, generating the requisite allosteric coupling, remains a rate-limiting step. Here, we describe an unbiased approach, termed domain-insertion profiling with DNA sequencing (DIP-seq), that combines the rapid creation of diverse libraries of potential SFPBs and high-throughput activity assays to identify functional biosensors. As a proof of concept, we construct an SFPB for the important regulatory sugar trehalose. DIP-seq analysis of a trehalose-binding-protein reveals allosteric hotspots for GFP insertion and results in high-dynamic range biosensors that function robustly in vivo. Taken together, DIP-seq simultaneously accelerates metabolite biosensor construction and provides a novel tool for interrogating protein allostery. PMID:27470466

  16. Rapid construction of metabolite biosensors using domain-insertion profiling.

    PubMed

    Nadler, Dana C; Morgan, Stacy-Anne; Flamholz, Avi; Kortright, Kaitlyn E; Savage, David F

    2016-01-01

    Single-fluorescent protein biosensors (SFPBs) are an important class of probes that enable the single-cell quantification of analytes in vivo. Despite advantages over other detection technologies, their use has been limited by the inherent challenges of their construction. Specifically, the rational design of green fluorescent protein (GFP) insertion into a ligand-binding domain, generating the requisite allosteric coupling, remains a rate-limiting step. Here, we describe an unbiased approach, termed domain-insertion profiling with DNA sequencing (DIP-seq), that combines the rapid creation of diverse libraries of potential SFPBs and high-throughput activity assays to identify functional biosensors. As a proof of concept, we construct an SFPB for the important regulatory sugar trehalose. DIP-seq analysis of a trehalose-binding-protein reveals allosteric hotspots for GFP insertion and results in high-dynamic range biosensors that function robustly in vivo. Taken together, DIP-seq simultaneously accelerates metabolite biosensor construction and provides a novel tool for interrogating protein allostery. PMID:27470466

  17. Thiol- and biotin-labeled probes for oligonucleotide quartz crystal microbalance biosensors of microalga alexandrium minutum.

    PubMed

    Lazerges, Mathieu; Perrot, Hubert; Rabehagasoa, Niriniony; Compère, Chantal

    2012-01-01

    Two quartz crystal microbalance oligonucleotide biosensors of a toxic microalga gene sequence (Alexandrium Minutum) have been designed. Grafting on a gold surface of 20-base thiol- or biotin-labeled probe, and selective hybridization with the complementary 20-base target, have been monitored in situ with a 27 MHz quartz crystal microbalance under controlled hydrodynamic conditions. The frequency of the set up is stable to within a few hertz, corresponding to the nanogram scale, for three hour experiments. DNA recognition by the two biosensors is efficient and selective. Hybridization kinetic curves indicate that the biosensor designed with the thiol-labeled probe is more sensitive, and that the biosensor designed with the biotin-labeled probe has a shorter time response and a higher hybridization efficiency. PMID:25585927

  18. Thiol- and Biotin-Labeled Probes for Oligonucleotide Quartz Crystal Microbalance Biosensors of Microalga Alexandrium Minutum

    PubMed Central

    Lazerges, Mathieu; Perrot, Hubert; Rabehagasoa, Niriniony; Compère, Chantal

    2012-01-01

    Two quartz crystal microbalance oligonucleotide biosensors of a toxic microalga gene sequence (Alexandrium Minutum) have been designed. Grafting on a gold surface of 20-base thiol- or biotin-labeled probe, and selective hybridization with the complementary 20-base target, have been monitored in situ with a 27 MHz quartz crystal microbalance under controlled hydrodynamic conditions. The frequency of the set up is stable to within a few hertz, corresponding to the nanogram scale, for three hour experiments. DNA recognition by the two biosensors is efficient and selective. Hybridization kinetic curves indicate that the biosensor designed with the thiol-labeled probe is more sensitive, and that the biosensor designed with the biotin-labeled probe has a shorter time response and a higher hybridization efficiency. PMID:25585927

  19. NANOSCALE BIOSENSORS IN ECOSYSTEM EXPOSURE RESEARCH

    EPA Science Inventory

    This powerpoint presentation presented information on nanoscale biosensors in ecosystem exposure research. The outline of the presentation is as follows: nanomaterials environmental exposure research; US agencies involved in nanosensor research; nanoscale LEDs in biosensors; nano...

  20. BIOSENSORS FOR ENVIRONMENTAL MONITORING: A REGULATORY PERSPECTIVE

    EPA Science Inventory

    Biosensors show the potential to complement laboratory-based analytical methods for environmental applications. Although biosensors for potential environmental-monitoring applications have been reported for a wide range of environmental pollutants, from a regulatory perspective, ...

  1. Gated Ion Channel-Based Biosensor Device

    NASA Astrophysics Data System (ADS)

    Separovic, Frances; Cornell, Bruce A.

    A biosensor device based on the ion channel gramicidin A (gA) incorporated into a bilayer membrane is described. This generic immunosensing device utilizes gA coupled to an antibody and assembled in a lipid membrane. The membrane is chemically tethered to a gold electrode, which reports on changes in the ionic conduction of the lipid bilayer. Binding of a target molecule in the bathing solution to the antibody causes the gramicidin channels to switch from predominantly conducting dimers to predominantly nonconducting monomers. Conventional a.c. impedance spectroscopy between the gold and a counter electrode in the bathing solution is used to measure changes in the ionic conductivity of the membrane. This approach permits the quantitative detection of a range of target species, including bacteria, proteins, toxins, DNA sequences, and drug molecules.

  2. Gold Nanoparticle Coated Silica Nanorods for Sensitive Visual Detection of microRNA on a Lateral Flow Strip Biosensor.

    PubMed

    Takalkar, Sunitha; Xu, Hui; Chen, Jiao; Baryeh, Kwaku; Qiu, Wanwei; Zhao, Julia X; Liu, And Guodong

    2016-01-01

    We present a rapid and highly sensitive approach for visual detection of microRNA (miRNA) using a gold nanoparticles coated silica nanorod label and lateral flow strip biosensor. Gold nanoparticles were decorated on the silica nanorod surface by a seeding and growth procedure. A single strand DNA probe was immobilized on the gold nanoparticles-silica nanorod surface by a self-assembling process, and the formed DNA-gold nanoparticles-silica nanorod conjugate was used to construct the lateral flow nucleic acid biosensor for detecting miRNA. The captured gold nanoparticles-silica nanorods by sandwich-type hybridization reactions (DNA-RNA-DNA) on the test zone of the lateral flow nucleic acid biosensor produced the characteristic color bands, enabling visual detection of miRNA. After systematic optimization, the new lateral flow nucleic acid biosensor was capable of detecting 10 pM of the miRNA target without instrumentation, which is six times lower than that obtained with the gold nanoparticle-based lateral flow nucleic acid biosensor. The gold nanoparticles coated silica nanorod thus provides a new and sensitive nanolabel for visual detection of biological molecules on the lateral flow biosensor. PMID:27302581

  3. Biosensor for metal analysis and speciation

    DOEpatents

    Aiken, Abigail M.; Peyton, Brent M.; Apel, William A.; Petersen, James N.

    2007-01-30

    A biosensor for metal analysis and speciation is disclosed. The biosensor comprises an electron carrier immobilized to a surface of an electrode and a layer of an immobilized enzyme adjacent to the electrode. The immobilized enzyme comprises an enzyme having biological activity inhibited by a metal to be detected by the biosensor.

  4. A third-order mode high frequency biosensor with atomic resolution.

    PubMed

    Cai, Hua-Lin; Yang, Yi; Chen, Xiao; Mohammad, Mohammad Ali; Ye, Tian-Xiang; Guo, Cang-Ran; Yi, Li-Ting; Zhou, Chang-Jian; Liu, Jing; Ren, Tian-Ling

    2015-09-15

    An atomic resolution ultra-high sensitivity surface acoustic wave (SAW) biosensor for DNA sequences and cells detection is proposed. Interdigitated transducers (IDTs) fabricated on LiNbO3 substrate achieve a high quality factor (Q) of over 4000 at a frequency of 6.4 GHz (third-order harmonic mode) using an optimized design and process. The biosensor shows excellent linear responses to target DNA in the range from 1 μg/ml to 1 ng/ml with a high sensitivity of 6.7 × 10(-16)g/cm(2)/Hz, hence the difference of a single hybridized DNA base can also be distinguished. With such a high mass resolution, the biosensor is capable of quantitative detection of living cancer cells. The frequency responses of single mouse mammary adenocarcinoma (EMT6) cell and mouse fibroblast (3T3) cell are studied. The interferences in the experiments show insignificant influence on the frequency shift, which verifies the high selectivity of the biosensor. The biosensor is also able to repeat the sensing ability after rough cleaning, therefore cost reduction is achieved from the recycling process in practical applications. The detection limit is defined from the noise analysis of the device, atomic resolution is realized according to the calculation, thereby initiating a potential tool for high-precision medical diagnoses and phenomena observation at the atomic-level. PMID:25913447

  5. 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

  6. Plants as Environmental Biosensors

    PubMed Central

    Ranatunga, Don Rufus A

    2006-01-01

    Plants are continuously exposed to a wide variety of perturbations including variation of temperature and/or light, mechanical forces, gravity, air and soil pollution, drought, deficiency or surplus of nutrients, attacks by insects and pathogens, etc., and hence, it is essential for all plants to have survival sensory mechanisms against such perturbations. Consequently, plants generate various types of intracellular and intercellular electrical signals mostly in the form of action and variation potentials in response to these environmental changes. However, over a long period, only certain plants with rapid and highly noticeable responses for environmental stresses have received much attention from plant scientists. Of particular interest to our recent studies on ultra fast action potential measurements in green plants, we discuss in this review the evidence supporting the foundation for utilizing green plants as fast biosensors for molecular recognition of the direction of light, monitoring the environment, and detecting the insect attacks as well as the effects of pesticides, defoliants, uncouplers, and heavy metal pollutants. PMID:19521490

  7. Biosensors for termite control

    NASA Astrophysics Data System (ADS)

    Farkhanda, M.

    2013-12-01

    Termites are major urban pests in Pakistan and cause damage to wooden structures and buildings. Termite management has two parts: prevention and control. The most difficult part of termite control is termite detection as most of them are subterranean in Pakistan and have tunneling habit.Throughout the world, chemical termiticides are going to be replaced by baits, microwave and sensor technology. Termite species are distinct biologically and have specific foraging behaviors. Termite Detection Radar, Moisture meter and Remote Thermal Sensor with Laser are available throughout the world. These can detect termites underground and use fewer chemicals than traditional methods. For wooden buildings, a termite sensor and an intrusion detection system for detecting termites are designed. A pair of electrodes is disposed inside the container. A pair of terminals is connected to these electrodes, these extend outside the container. Termites are detected by a change of conductivity between the electrodes, when termites are detected a warning device generates a warning signal. In Pakistan, there is dire need to develop such biosensoring devices locally, then apply control methods that would save money and protect the environment.

  8. Fluidics cube for biosensor miniaturization

    NASA Technical Reports Server (NTRS)

    Dodson, J. M.; Feldstein, M. J.; Leatzow, D. M.; Flack, L. K.; Golden, J. P.; Ligler, F. S.

    2001-01-01

    To create a small, portable, fully automated biosensor, a compact means of fluid handling is required. We designed, manufactured, and tested a "fluidics cube" for such a purpose. This cube, made of thermoplastic, contains reservoirs and channels for liquid samples and reagents and operates without the use of any internal valves or meters; it is a passive fluid circuit that relies on pressure relief vents to control fluid movement. We demonstrate the ability of pressure relief vents to control fluid movement and show how to simply manufacture or modify the cube. Combined with the planar array biosensor developed at the Naval Research Laboratory, it brings us one step closer to realizing our goal of a handheld biosensor capable of analyzing multiple samples for multiple analytes.

  9. Enzymatic Polymerization on DNA Modified Gold Nanowire for Label-Free Detection of Pathogen DNA

    PubMed Central

    Jeong, Jaepil; Kim, Hyejin; Lee, Jong Bum

    2015-01-01

    This paper presents a label-free biosensor for the detection of single-stranded pathogen DNA through the target-enhanced gelation between gold nanowires (AuNW) and the primer DNAs branched on AuNW. The target DNA enables circularization of the linear DNA template, and the primer DNA is elongated continuously via rolling circle amplification. As a result, in the presence of the target DNA, a macroscopic hydrogel was fabricated by the entanglement of the elongated DNA with AuNWs as a scaffold fiber for effective gelation. In contrast, very small separate particles were generated in the absence of the target DNA. This label-free biosensor might be a promising tool for the detection of pathogen DNAs without any devices for further analysis. Moreover, the biosensor based on the weaving of AuNW and DNAs suggests a novel direction for the applications of AuNWs in biological engineering. PMID:26084045

  10. Modelling a Peroxidase-based Optical Biosensor

    PubMed Central

    Baronas, Romas; Gaidamauskaite, Evelina; Kulys, Juozas

    2007-01-01

    The response of a peroxidase-based optical biosensor was modelled digitally. A mathematical model of the optical biosensor is based on a system of non-linear reaction-diffusion equations. The modelling biosensor comprises two compartments, an enzyme layer and an outer diffusion layer. The digital simulation was carried out using finite difference technique. The influence of the substrate concentration as well as of the thickness of both the enzyme and diffusion layers on the biosensor response was investigated. Calculations showed complex kinetics of the biosensor response, especially at low concentrations of the peroxidase and of the hydrogen peroxide.

  11. Biosensors for hepatitis B virus detection.

    PubMed

    Yao, Chun-Yan; Fu, Wei-Ling

    2014-09-21

    A biosensor is an analytical device used for the detection of analytes, which combines a biological component with a physicochemical detector. Recently, an increasing number of biosensors have been used in clinical research, for example, the blood glucose biosensor. This review focuses on the current state of biosensor research with respect to efficient, specific and rapid detection of hepatitis B virus (HBV). The biosensors developed based on different techniques, including optical methods (e.g., surface plasmon resonance), acoustic wave technologies (e.g., quartz crystal microbalance), electrochemistry (amperometry, voltammetry and impedance) and novel nanotechnology, are also discussed. PMID:25253948

  12. Adapting selected nucleic acid ligands (aptamers) to biosensors.

    PubMed

    Potyrailo, R A; Conrad, R C; Ellington, A D; Hieftje, G M

    1998-08-15

    A flexible biosensor has been developed that utilizes immobilized nucleic acid aptamers to specifically detect free nonlabeled non-nucleic acid targets such as proteins. In a model system, an anti-thrombin DNA aptamer was fluorescently labeled and covalently attached to a glass support. Thrombin in solution was selectively detected by following changes in the evanescent-wave-induced fluorescence anisotropy of the immobilized aptamer. The new biosensor can detect as little as 0.7 amol of thrombin in a 140-pL interrogated volume, has a dynamic range of 3 orders of magnitude, has an inter-sensing-element measurement precision of better than 4% RSD over the range 0-200 nM, and requires less than 10 min for sample analysis. The aptamer-sensor format is generalizable and should allow sensitive, selective, and fast determination of a wide range of analytes. PMID:9726167

  13. Biosensors in Clinical Practice: Focus on Oncohematology

    PubMed Central

    Fracchiolla, Nicola S.; Artuso, Silvia; Cortelezzi, Agostino

    2013-01-01

    Biosensors are devices that are capable of detecting specific biological analytes and converting their presence or concentration into some electrical, thermal, optical or other signal that can be easily analysed. The first biosensor was designed by Clark and Lyons in 1962 as a means of measuring glucose. Since then, much progress has been made and the applications of biosensors are today potentially boundless. This review is limited to their clinical applications, particularly in the field of oncohematology. Biosensors have recently been developed in order to improve the diagnosis and treatment of patients affected by hematological malignancies, such as the biosensor for assessing the in vitro pre-treatment efficacy of cytarabine in acute myeloid leukemia, and the fluorescence resonance energy transfer-based biosensor for assessing the efficacy of imatinib in chronic myeloid leukemia. The review also considers the challenges and future perspectives of biosensors in clinical practice. PMID:23673681

  14. 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

  15. Smart built-in test

    NASA Technical Reports Server (NTRS)

    Richards, Dale W.

    1990-01-01

    The work which built-in test (BIT) is asked to perform in today's electronic systems increases with every insertion of new technology or introduction of tighter performance criteria. Yet the basic purpose remains unchanged -- to determine with high confidence the operational capability of that equipment. Achievement of this level of BIT performance requires the management and assimilation of a large amount of data, both realtime and historical. Smart BIT has taken advantage of advanced techniques from the field of artificial intelligence (AI) in order to meet these demands. The Smart BIT approach enhances traditional functional BIT by utilizing AI techniques to incorporate environmental stress data, temporal BIT information and maintenance data, and realtime BIT reports into an integrated test methodology for increased BIT effectiveness and confidence levels. Future research in this area will incorporate onboard fault-logging of BIT output, stress data and Smart BIT decision criteria in support of a singular, integrated and complete test and maintenance capability. The state of this research is described along with a discussion of directions for future development.

  16. A luminescent nisin biosensor

    NASA Astrophysics Data System (ADS)

    Immonen, Nina; Karp, Matti

    2006-02-01

    Nisin is a lantibiotic, an antibacterial peptide produced by certain Lactococcus lactis strains that kills or inhibits the growth of other bacteria. Nisin is widely used as a food preservative, and its long-time use suggests that it can be generally regarded as safe. We have developed a method for determining the amount of nisin in food samples that is based on luminescent biosensor bacteria. Bacterial luciferase operon luxABCDE was inserted into plasmid pNZ8048, and the construct was transformed by electroporation into Lc. lactis strain NZ9800, whose ability to produce nisin has been erased by deletion of the gene nisA. The operon luxABCDE has been modified to be functional in gram-positive bacteria to confer a bioluminescent phenotype without the requirement of adding an exogenous substrate. In the plasmid pNZ8048, the operon was placed under control of the nisin-inducible nisA promoter. The chromosomal nisRK genes of Lc. lactis NZ9800 allow it to sense nisin in the environment and relay this signal via signal transduction proteins NisK and NisR to initiate transcription from nisA promoter. In the case of our sensor bacteria, this leads to production of luciferase and, thus, luminescence that can be directly measured from living bacteria. Luminescence can be detected as early as within minutes of induction. The nisin assay described here provides a detection limit in the sub-picogram level per ml, and a linear area between 1 - 1000 pg/ml. The sensitivity of this assay exceeds the performance of all previously published methods.

  17. Electrical Percolation Based Biosensors

    PubMed Central

    Bruck, Hugh Alan; Yang, Minghui; Kostov, Yordan; Rasooly, Avraham

    2013-01-01

    A new approach to label free biosensing has been developed based on the principle of “electrical percolation”. In electrical percolation, long-range electrical connectivity is formed in randomly oriented and distributed systems of discrete elements. By applying this principle to biological interactions, it is possible to measure biological components both directly and electronically. The main element for electrical percolation biosensor is the biological semiconductor (BSC) which is a multi-layer 3-D carbon nanotube-antibody network. In the BSC, molecular interactions, such as binding of antigens to the antibodies, disrupt the network continuity causing increased resistance of the network. BSCs can be fabricated by immobilizing conducting elements, such as pre-functionalized single-walled carbon nanotubes (SWNTs)-antibody complex, directly onto a substrate, such as a Poly(methyl methacrylate) (PMMA) surface (also known as plexi-glass or Acrylic). BSCs have been demonstrated for direct (label-free) electronic measurements of antibody-antigen binding using SWNTs. If the concentration of the SWNT network is slightly above the electrical percolation threshold, then binding of a specific antigen to the pre-functionalized SWNT dramatically increases the electrical resistance due to changes in the tunneling between the SWNTs. Using anti-Staphylococcal enterotoxin B (SEB) IgG as a “gate” and SEB as an “actuator”, it was demonstrated that the BSC was able to detect SEB at concentrations of 1 ng/ml. Based on this concept, an automated configuration for BSCs is described here that enables real time continuous detection. The new BSC configuration may permit assembly of multiple sensors on the same chip to create “Biological Central Processing Units (CPUs)” with multiple biological elements, capable of processing and sorting out information on multiple analytes simultaneously. PMID:24041756

  18. DESIGN OF INTEGRATING WAVEGUIDE BIOSENSOR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Integrating Waveguide Biosensor allows for rapid and sensitive detection of pathogenic agents, cells and proteins via immunoassay or PCR products. The analytes are captured on the surface of the waveguide and then tagged with fluorescent labels. The waveguides are illuminated by excitation light...

  19. Improved Ion-Channel Biosensors

    NASA Technical Reports Server (NTRS)

    Nadeau, Jay; White, Victor; Dougherty, Dennis; Maurer, Joshua

    2004-01-01

    An effort is underway to develop improved biosensors of a type based on ion channels in biomimetic membranes. These sensors are microfabricated from silicon and other materials compatible with silicon. As described, these sensors offer a number of advantages over prior sensors of this type.

  20. A high content assay for biosensor validation and for examining stimuli that affect biosensor activity

    PubMed Central

    Slattery, Scott D.; Hahn, Klaus M.

    2015-01-01

    Biosensors are valuable tools used to monitor many different protein behaviors in vivo. Demand for new biosensors is high, but their development and characterization can be difficult. During biosensor design, it is necessary to evaluate the effects of different biosensor structures on specificity, brightness, and fluorescence responses. By co-expressing the biosensor with upstream proteins that either stimulate or inhibit the activity reported by the biosensor, one can determine the difference between the biosensor’s maximally activated and inactivated state, and examine response to specific proteins. This involves considerable labor and expense, as expression conditions must be optimized to saturate the biosensor with the regulator, and multiple replicates and controls are required. We describe here a protocol for biosensor validation in a 96-well plate format using an automated microscope. This protocol produces dose-response curves, enables efficient examination of many parameters, and unlike cell suspension assays allows visual inspection (eg for cell health and biosensor or regulator localization). Optimization of single chain and dual chain Rho GTPase biosensors is addressed, but the assay is applicable to any biosensor that can be expressed or otherwise loaded in adherent cells. The assay can also be used for purposes other than biosensor validation, using a well-characterized biosensor as a readout for variations in upstream molecules. PMID:25447074

  1. Tiny Medicine: Nanomaterial-Based Biosensors

    PubMed Central

    Yun, Yeo-Heung; Eteshola, Edward; Bhattacharya, Amit; Dong, Zhongyun; Shim, Joon-Sub; Conforti, Laura; Kim, Dogyoon; Schulz, Mark J.; Ahn, Chong H.; Watts, Nelson

    2009-01-01

    Tiny medicine refers to the development of small easy to use devices that can help in the early diagnosis and treatment of disease. Early diagnosis is the key to successfully treating many diseases. Nanomaterial-based biosensors utilize the unique properties of biological and physical nanomaterials to recognize a target molecule and effect transduction of an electronic signal. In general, the advantages of nanomaterial-based biosensors are fast response, small size, high sensitivity, and portability compared to existing large electrodes and sensors. Systems integration is the core technology that enables tiny medicine. Integration of nanomaterials, microfluidics, automatic samplers, and transduction devices on a single chip provides many advantages for point of care devices such as biosensors. Biosensors are also being used as new analytical tools to study medicine. Thus this paper reviews how nanomaterials can be used to build biosensors and how these biosensors can help now and in the future to detect disease and monitor therapies. PMID:22291565

  2. A biosensor for organoarsenical herbicides and growth promoters

    PubMed Central

    Chen, Jian; Sun, Samio; Li, Chen-Zhong; Zhu, Yong-Guan; Rosen, Barry P.

    2014-01-01

    The toxic metalloid arsenic is widely distributed in food, water, and soil. While inorganic arsenic enters the environment primarily from geochemical sources, methylarsenicals either result from microbial biotransformation of inorganic arsenic or are introduced anthropogenically. Methylarsenicals such as monosodium methylarsonic acid (MSMA) have been extensively utilized as herbicides, and aromatic arsenicals such as roxarsone (Rox) are used as growth promoters for poultry and swine. Organoarsenicals are degraded to inorganic arsenic. The toxicological effects of arsenicals depend on their oxidation state, chemical composition, and bioavailability. Here we report that the active forms are the trivalent arsenic-containing species. We constructed a whole-cell biosensor utilizing a modified ArsR repressor that is highly selective toward trivalent methyl and aromatic arsenicals, with essentially no response to inorganic arsenic. The biosensor was adapted for in vitro detection of organoarsenicals using fluorescence anisotropy of ArsR-DNA interactions. It detects bacterial biomethylation of inorganic arsenite both in vivo and in vitro with detection limits of 10−7 M and linearity to 10−6 M for phenylarsenite and 5×10−6 M for methylarsenite. The biosensor detects reduced forms of MSMA and roxarsone and offers a practical, low cost method for detecting activate forms and breakdown products of organoarsenical herbicides and growth promoters. PMID:24359149

  3. Recent Development in Optical Fiber Biosensors

    PubMed Central

    Bosch, María Espinosa; Sánchez, Antonio Jesús Ruiz; Rojas, Fuensanta Sánchez; Ojeda, Catalina Bosch

    2007-01-01

    Remarkable developments can be seen in the field of optical fibre biosensors in the last decade. More sensors for specific analytes have been reported, novel sensing chemistries or transduction principles have been introduced, and applications in various analytical fields have been realised. This review consists of papers mainly reported in the last decade and presents about applications of optical fiber biosensors. Discussions on the trends in optical fiber biosensor applications in real samples are enumerated.

  4. Artificial DNA and surface plasmon resonance

    PubMed Central

    D'Agata, Roberta; Spoto, Giuseppe

    2012-01-01

    The combined use of surface plasmon resonance (SPR) and modified or mimic oligonucleotides have expanded diagnostic capabilities of SPR-based biosensors and have allowed detailed studies of molecular recognition processes. This review summarizes the most significant advances made in this area over the past 15 years.   Functional and conformationally restricted DNA analogs (e.g., aptamers and PNAs) when used as components of SPR biosensors contribute to enhance the biosensor sensitivity and selectivity. At the same time, the SPR technology brings advantages that allows forbetter exploration of underlying properties of non-natural nucleic acid structures such us DNAzymes, LNA and HNA. PMID:22821257

  5. Survey of the 1998 optical biosensor literature.

    PubMed

    Myszka, D G

    1999-01-01

    The utilization of optical biosensors to study molecular interactions continues to expand. In 1998, 384 articles relating to the use of commercial biosensors were published in 130 different journals. While significant strides in new applications and methodology were made, a majority of the biosensor literature is of rather poor quality. Basic information about experimental conditions is often not presented and many publications fail to display the experimental data, bringing into question the credibility of the results. This review provides suggestions on how to collect, analyze and report biosensor data. PMID:10611648

  6. Zinc oxide interdigitated electrode for biosensor application

    NASA Astrophysics Data System (ADS)

    Sin L., L.; Arshad, M. K. Md.; Fathil, M. F. M.; Adzhri, R.; M. Nuzaihan M., N.; Ruslinda, A. R.; Gopinath, Subash C. B.; Hashim, U.

    2016-07-01

    In biosensors, zinc oxide (ZnO) thin film plays a crucial role in term of stability, sensitivity, biocompatibility and low cost. Interdigitated electrode (IDE) design is one of the device architecture in biosensor for label free, stability and sensitivity. In this paper, we discuss the fabrication of zinc oxide deposited on the IDE as a transducer for sensing of biomolecule. The formation of APTES had increase the performance of the surface functionalization..Furthermore we extend the discuss on the surface functionalization process which is utilized for probe attachment onto the surface of biosensor through surface immobilization process, thus enables the sensing of biomolecules for biosensor application.

  7. Portable evanescent wave fiber biosensor for highly sensitive detection of Shigella

    NASA Astrophysics Data System (ADS)

    Xiao, Rui; Rong, Zhen; Long, Feng; Liu, Qiqi

    2014-11-01

    A portable evanescent wave fiber biosensor was developed to achieve the rapid and highly sensitive detection of Shigella. In this study, a DNA probe was covalently immobilized onto fiber-optic biosensors that can hybridize with a fluorescently labeled complementary DNA. The sensitivity of detection for synthesized oligonucleotides can reach 10-10 M. The surface of the sensor can be regenerated with 0.5% sodium dodecyl sulfate solution (pH 1.9) for over 30 times without significant deterioration of performance. The total analysis time for a single sample, including the time for measurement and surface regeneration, was less than 6 min. We employed real-time polymerase chain reaction (PCR) and compared the results of both methods to investigate the actual Shigella DNA detection capability of the fiber-optic biosensor. The fiber-optic biosensor could detect as low as 102 colony-forming unit/mL Shigella. This finding was comparable with that by real-time PCR, which suggests that this method is a potential alternative to existing detection methods.

  8. Biosensor based on magnetostrictive microcantilever

    NASA Astrophysics Data System (ADS)

    Li, Suiqiong; Orona, Lisa; Li, Zhimin; Cheng, Z.-Y.

    2006-02-01

    Magnetostrictive microcantilever (MSMC) as remote biosensor platform is reported. The mass sensitivity of the MSMCs is simulated and compared with the other microcantilevers. MSMCs with a thickness of 30-35μm and different lengths and widths were fabricated from the magnetostrictive metal glass coated with a copper layer by sputtering. The resonance behavior of the MSMCs was experimentally determined. It is experimentally found that the MSMCs work well in either air or liquid. For MSMCs operated in air, a Q value of more than 500 was obtained. For MSMCs operated in water, the Q value reaches more than 30. The application of a MSMC as a biosensor platform is demonstrated by in situ detection of the yeast cells in water using the MSMC sensor.

  9. Optical biosensors for environmental monitoring

    SciTech Connect

    Tamiya, Eiichi

    1996-12-31

    Environmental assessment is important to evaluate the overall health and ecological impact of domestic and industrial wastes. Biosensors are kinds of analytical devices which consist of biomaterials and transducers. Photoluminescence of recombinant E. coli containing lux related genes were used as indicators of environmental pollutions. This paper deals with sensitive and rapid optical sensing systems for monitoring BOD (Biochemical Oxygen Demand), toxic compounds and mutagens.

  10. Alginate cryogel based glucose biosensor

    NASA Astrophysics Data System (ADS)

    Fatoni, Amin; Windy Dwiasi, Dian; Hermawan, Dadan

    2016-02-01

    Cryogel is macroporous structure provides a large surface area for biomolecule immobilization. In this work, an alginate cryogel based biosensor was developed to detect glucose. The cryogel was prepared using alginate cross-linked by calcium chloride under sub-zero temperature. This porous structure was growth in a 100 μL micropipette tip with a glucose oxidase enzyme entrapped inside the cryogel. The glucose detection was based on the colour change of redox indicator, potassium permanganate, by the hydrogen peroxide resulted from the conversion of glucose. The result showed a porous structure of alginate cryogel with pores diameter of 20-50 μm. The developed glucose biosensor was showed a linear response in the glucose detection from 1.0 to 5.0 mM with a regression of y = 0.01x+0.02 and R2 of 0.994. Furthermore, the glucose biosensor was showed a high operational stability up to 10 times of uninterrupted glucose detections.

  11. Biosensors based on nanomechanical systems.

    PubMed

    Tamayo, Javier; Kosaka, Priscila M; Ruz, José J; San Paulo, Álvaro; Calleja, Montserrat

    2013-02-01

    The advances in micro- and nanofabrication technologies enable the preparation of increasingly smaller mechanical transducers capable of detecting the forces, motion, mechanical properties and masses that emerge in biomolecular interactions and fundamental biological processes. Thus, biosensors based on nanomechanical systems have gained considerable relevance in the last decade. This review provides insight into the mechanical phenomena that occur in suspended mechanical structures when either biological adsorption or interactions take place on their surface. This review guides the reader through the parameters that change as a consequence of biomolecular adsorption: mass, surface stress, effective Young's modulus and viscoelasticity. The mathematical background needed to correctly interpret the output signals from nanomechanical biosensors is also outlined here. Other practical issues reviewed are the immobilization of biomolecular receptors on the surface of nanomechanical systems and methods to attain that in large arrays of sensors. We then describe some relevant realizations of biosensor devices based on nanomechanical systems that harness some of the mechanical effects cited above. We finally discuss the intrinsic detection limits of the devices and the limitation that arises from non-specific adsorption. PMID:23152052

  12. Biosensor of endotoxin and sepsis

    NASA Astrophysics Data System (ADS)

    Shao, Yang; Wang, Xiang; Wu, Xi; Gao, Wei; He, Qing-hua; Cai, Shaoxi

    2001-09-01

    To investigate the relation between biosensor of endotoxin and endotoxin of plasma in sepsis. Method: biosensor of endotoxin was designed with technology of quartz crystal microbalance bioaffinity sensor ligand of endotoxin were immobilized by protein A conjugate. When a sample soliton of plasma containing endotoxin 0.01, 0.03, 0.06, 0.1, 0.5, 1.0Eu, treated with perchloric acid and injected into slot of quartz crystal surface respectively, the ligand was released from the surface of quartz crystal to form a more stable complex with endotoxin in solution. The endotoxin concentration corresponded to the weight change on the crystal surface, and caused change of frequency that occurred when desorbed. The result was biosensor of endotoxin might detect endotoxin of plasma in sepsis, measurements range between 0.05Eu and 0.5Eu in the stop flow mode, measurement range between 0.1Eu and 1Eu in the flow mode. The sensor of endotoxin could detect the endotoxin of plasm rapidly, and use for detection sepsis in clinically.

  13. Four-channel label-free photonic crystal biosensor using nanocavity resonators

    NASA Astrophysics Data System (ADS)

    Olyaee, Saeed; Najafgholinezhad, Samira; Alipour Banaei, Hamed

    2013-09-01

    In this paper, we design and characterize a novel small size four-channel biosensor based on the two-dimensional photonic crystal with introducing waveguides and nano-cavities in the hexagonal lattice of air pores in the silicon slab. By removing a group of air pores, waveguides are achieved, and nano-cavities are shaped by modifying the radius of air pores. Highly parallel operation of this biosensor due to the special architecture is the capability of the designed structure. The biomaterials which are suspended in a liquid medium inside nano-cavities cause effective refractive index changes which lead to the resonant wavelength shift in the output terminal. According to results, with increasing the refractive index of nano-cavities, resonant wavelengths shifts to longer values. For biochemical sensing like DNA molecule and protein and for the refractive index detection, this novel designed biosensor can be utilized.

  14. Post-Translational Modification of Bionanoparticles as a Modular Platform for Biosensor Assembly.

    PubMed

    Sun, Qing; Chen, Qi; Blackstock, Daniel; Chen, Wilfred

    2015-08-25

    Context driven biosensor assembly with modular targeting and detection moieties is gaining significant attentions. Although protein-based nanoparticles have emerged as an excellent platform for biosensor assembly, current strategies of decorating bionanoparticles with targeting and detection moieties often suffer from unfavorable spacing and orientation as well as bionanoparticle aggregation. Herein, we report a highly modular post-translational modification approach for biosensor assembly based on sortase A-mediated ligation. This approach enables the simultaneous modifications of the Bacillus stearothermophilus E2 nanoparticles with different functional moieties for antibody, enzyme, DNA aptamer, and dye decoration. The resulting easy-purification platform offers a high degree of targeting and detection modularity with signal amplification. This flexibility is demonstrated for the detection of both immobilized antigens and cancer cells. PMID:26235232

  15. MWCNTs based high sensitive lateral flow strip biosensor for rapid determination of aqueous mercury ions.

    PubMed

    Yao, Li; Teng, Jun; Zhu, Mengya; Zheng, Lei; Zhong, Youhao; Liu, Guodong; Xue, Feng; Chen, Wei

    2016-11-15

    Here, we describe a disposable multi-walled carbon nanotubes (MWCNTs) labeled nucleic acid lateral flow strip biosensor for rapid and sensitive detection of aqueous mercury ions (Hg(2+)). Unlike the conventional colloidal gold nanoparticle based strip biosensors, the carboxylated MWCNTs were selected as the labeling substrate because of its high specific surface area for immobilization of recognition probes, improved stability and enhanced detection sensitivity of the strip biosensor. Combining the sandwich-type of T-Hg(2+)-T recognition mechanism with the optical properties of MWCNTs on lateral flow strip, optical black bands were observed on the lateral flow strips. Parameters (such as membrane category, the MWCNTs concentration, the amount of MWCNT-DNA probe, and the volume of the test probe) that govern the sensitivity and reproducibility of the sensor were optimized. The response of the optimized biosensor was highly linear over the range of 0.05-1ppb target Hg(2+), and the detection threshold was estimated at 0.05 ppb within a 15-min assay time. The sensitivity was 10-fold higher than the conventional colloidal gold based strip biosensor. More importantly, the stability of the sensor was also greatly improved with the usage of MWCNTs as the labeling. PMID:27183284

  16. Built Environment Education in Art Education.

    ERIC Educational Resources Information Center

    Guilfoil, Joanne K., Ed.; Sandler, Alan R., Ed.

    This anthology brings the study of the built environment, its design, social and cultural functions, and the criticism thereof into focus. Following a preface and introduction, 22 essays are organized in three parts. Part 1 includes: (1) "Landscape Art and the Role of the Natural Environment in Built Environment Education" (Heather Anderson); (2)…

  17. Nanomaterials based biosensors for cancer biomarker detection

    NASA Astrophysics Data System (ADS)

    Malhotra, Bansi D.; Kumar, Saurabh; Mouli Pandey, Chandra

    2016-04-01

    Biosensors have enormous potential to contribute to the evolution of new molecular diagnostic techniques for patients suffering with cancerous diseases. A major obstacle preventing faster development of biosensors pertains to the fact that cancer is a highly complex set of diseases. The oncologists currently rely on a few biomarkers and histological characterization of tumors. Some of the signatures include epigenetic and genetic markers, protein profiles, changes in gene expression, and post-translational modifications of proteins. These molecular signatures offer new opportunities for development of biosensors for cancer detection. In this context, conducting paper has recently been found to play an important role towards the fabrication of a biosensor for cancer biomarker detection. In this paper we will focus on results of some of the recent studies obtained in our laboratories relating to fabrication and application of nanomaterial modified paper based biosensors for cancer biomarker detection.

  18. A creatinine biosensor based on admittance measurement

    NASA Astrophysics Data System (ADS)

    Ching, Congo Tak-Shing; Sun, Tai-Ping; Jheng, Deng-Yun; Tsai, Hou-Wei; Shieh, Hsiu-Li

    2015-08-01

    Regular check of blood creatinine level is very important as it is a measurement of renal function. Therefore, the objective of this study is to develop a simple and reliable creatinine biosensor based on admittance measurement for precise determination of creatinine. The creatinine biosensor was fabricated with creatinine deiminase immobilized on screen-printed carbon electrodes. Admittance measurement at a specific frequency ranges (22.80 - 84.71 Hz) showed that the biosensor has an excellent linear (r2 > 0.95) response range (50 - 250 uM), which covers the normal physiological and pathological ranges of blood creatinine levels. Intraclass correlation coefficient (ICC) showed that the biosensor has excellent reliability and validity (ICC = 0.98). In conclusion, a simple and reliable creatinine biosensor was developed and it is capable of precisely determining blood creatinine levels in both the normal physiological and pathological ranges.

  19. Biosensoric potential of microbial fuel cells.

    PubMed

    Schneider, György; Kovács, Tamás; Rákhely, Gábor; Czeller, Miklós

    2016-08-01

    Recent progress in microbial fuel cell (MFC) technology has highlighted the potential of these devices to be used as biosensors. The advantages of MFC-based biosensors are that they are phenotypic and can function in either assay- or flow-through formats. These features make them appropriate for contiguous on-line monitoring in laboratories and for in-field applications. The selectivity of an MFC biosensor depends on the applied microorganisms in the anodic compartment where electron transfer (ET) between the artificial surface (anode) and bacterium occurs. This process strongly determines the internal resistance of the sensoric system and thus influences signal outcome and response time. Despite their beneficial characteristics, the number of MFC-based biosensoric applications has been limited until now. The aim of this mini-review is to turn attention to the biosensoric potential of MFCs by summarizing ET mechanisms on which recently established and future sensoric devices are based. PMID:27401925

  20. TIGER: the universal biosensor

    NASA Astrophysics Data System (ADS)

    Hofstadler, Steven A.; Sampath, Rangarajan; Blyn, Lawrence B.; Eshoo, Mark W.; Hall, Thomas A.; Jiang, Yun; Drader, Jared J.; Hannis, James C.; Sannes-Lowery, Kristin A.; Cummins, Lendell L.; Libby, Brian; Walcott, Demetrius J.; Schink, Amy; Massire, Christian; Ranken, Raymond; Gutierrez, Jose; Manalili, Sheri; Ivy, Cristina; Melton, Rachael; Levene, Harold; Barrett-Wilt, Greg; Li, Feng; Zapp, Vanessa; White, Neill; Samant, Vivek; McNeil, John A.; Knize, Duane; Robbins, David; Rudnick, Karl; Desai, Anjali; Moradi, Emily; Ecker, David J.

    2005-03-01

    In this work, we describe a strategy for the detection and characterization of microorganisms associated with a potential biological warfare attack or a natural outbreak of an emerging infectious disease. This approach, termed TIGER (Triangulation Identification for the Genetic Evaluation of Risks), relies on mass spectrometry-derived base composition signatures obtained from PCR amplification of broadly conserved regions of the microbial genome(s) in a sample. The sample can be derived from air filtration devices, clinical samples, or other sources. Core to this approach are "intelligent PCR primers" that target broadly conserved regions of microbial genomes that flank variable regions. This approach requires that high-performance mass measurements be made on PCR products in the 80-140 bp size range in a high-throughput, robust modality. As will be demonstrated, the concept is equally applicable to bacteria and viruses and could be further applied to fungi and protozoa. In addition to describing the fundamental strategy of this approach, several specific examples of TIGER are presented that illustrate the impact this approach could have on the way biological weapons attacks are detected and the way that the etiologies of infectious diseases are determined. The first example illustrates how any bacterial species might be identified, using Bacillus anthracis as the test agent. The second example demonstrates how DNA-genome viruses are identified using five members of Poxviridae family, whose members includes Variola virus, the agent responsible for smallpox. The third example demonstrates how RNA-genome viruses are identified using the Alphaviruses (VEE, WEE, and EEE) as representative examples. These examples illustrate how the TIGER technology can be applied to create a universal identification strategy for all pathogens, including those that infect humans, livestock, and plants.

  1. Quantum dot-based microfluidic biosensor for cancer detection

    NASA Astrophysics Data System (ADS)

    Ghrera, Aditya Sharma; Pandey, Chandra Mouli; Ali, Md. Azahar; Malhotra, Bansi Dhar

    2015-05-01

    We report results of the studies relating to fabrication of an impedimetric microfluidic-based nucleic acid sensor for quantification of DNA sequences specific to chronic myelogenous leukemia (CML). The sensor chip is prepared by patterning an indium-tin-oxide (ITO) coated glass substrate via wet chemical etching method followed by sealing with polydimethylsiloxane (PDMS) microchannel for fluid control. The fabricated microfluidic chip comprising of a patterned ITO substrate is modified by depositing cadmium selenide quantum dots (QCdSe) via Langmuir-Blodgett technique. Further, the QCdSe surface has been functionalized with specific DNA probe for CML detection. The probe DNA functionalized QCdSe integrated miniaturized system has been used to monitor target complementary DNA concentration by measuring the interfacial charge transfer resistance via hybridization. The presence of complementary DNA in buffer solution significantly results in decreased electro-conductivity of the interface due to presence of a charge barrier for transport of the redox probe ions. The microfluidic DNA biosensor exhibits improved linearity in the concentration range of 10-15 M to 10-11 M.

  2. Quantum dot-based microfluidic biosensor for cancer detection

    SciTech Connect

    Ghrera, Aditya Sharma; Pandey, Chandra Mouli; Ali, Md. Azahar; Malhotra, Bansi Dhar

    2015-05-11

    We report results of the studies relating to fabrication of an impedimetric microfluidic–based nucleic acid sensor for quantification of DNA sequences specific to chronic myelogenous leukemia (CML). The sensor chip is prepared by patterning an indium–tin–oxide (ITO) coated glass substrate via wet chemical etching method followed by sealing with polydimethylsiloxane (PDMS) microchannel for fluid control. The fabricated microfluidic chip comprising of a patterned ITO substrate is modified by depositing cadmium selenide quantum dots (QCdSe) via Langmuir–Blodgett technique. Further, the QCdSe surface has been functionalized with specific DNA probe for CML detection. The probe DNA functionalized QCdSe integrated miniaturized system has been used to monitor target complementary DNA concentration by measuring the interfacial charge transfer resistance via hybridization. The presence of complementary DNA in buffer solution significantly results in decreased electro-conductivity of the interface due to presence of a charge barrier for transport of the redox probe ions. The microfluidic DNA biosensor exhibits improved linearity in the concentration range of 10{sup −15} M to 10{sup −11} M.

  3. Graphene oxide-based electrochemical label-free detection of glycoproteins down to aM level using a lectin biosensor.

    PubMed

    Klukova, L; Filip, J; Belicky, S; Vikartovska, A; Tkac, J

    2016-07-21

    A label-free ultrasensitive impedimetric biosensor with lectin immobilised on graphene oxide (GO) for the detection of glycoproteins from 1 aM is shown here. This is the first time a functional lectin biosensor with lectin directly immobilised on a graphene-based interface without any polymer modifier has been described. The study also shows that hydrophilic oxidative debris present on GO has a beneficial effect on the sensitivity of (8.46 ± 0.20)% per decade for the lectin biosensor compared to the sensitivity of (4.52 ± 0.23)% per decade for the lectin biosensor built up from GO with the oxidative debris washed out. PMID:27277703

  4. Self-assembled biosensor with universal reporter and dual-quenchers for detection of unlabelled nucleic acids.

    PubMed

    Huang, Liming; Aryal, Gyan H; Tam-Chang, Suk-Wah; Publicover, Nelson G; Hunter, Kenneth W

    2016-02-21

    A novel biosensor with universal reporter and dual quenchers was developed for rapid, sensitive, selective, and inexpensive detection of unlabelled nucleic acids. The biosensor is based on a single-strand DNA stem-loop motif with an extended universal reporter-binding region, a G-base rich stem region, and a universal address-binding region. The self-assembly of these stem-loop probes with fluorescence labeled universal reporter and a universal address region conjugated to gold nanoparticles forms the basis of a biosensor for DNA or microRNA targets in solution. The introduction of dual quenchers (G-base quenching and gold surface plasmon resonance-induced quenching) significantly reduces the fluorescence background to as low as 12% of its original fluorescence intensity and hence enhances the detection limit to 0.01 picomoles without signal ampilication. PMID:26757447

  5. Meeting current public health needs: optical biosensors for pathogen detection and analysis

    NASA Astrophysics Data System (ADS)

    Yang, Minghui; Sapsford, Kim E.; Sergeev, Nikolay; Sun, Steven; Rasooly, Avraham

    2009-02-01

    Pathogen detection and analysis is critical for medicine, food safety, agriculture, public health and biosecurity. Many current microbial detection approaches are based on century-old culturing methods which, while reliable, are slow, provide relatively little information about the pathogens and are not adaptable to high throughput operations. Optical biodetection represents a potential alternative. Most ELISA and chromatography systems are based on optical methods that are also used for analysis of molecular interactions, such as DNA hybridization and protein-protein interactions (e.g. microarrays or SPR biosensors). Various optical biosensor platforms have been developed that have many of the characteristics essential for modern pathogen molecular analysis including sensitivity, speed of analysis, multi-channel capability, relative simplicity and low cost. Here we provide several examples of the use of optical biosensor technology for pathogen detection and analysis including high throughput DNA microarray analysis, SPR-based rapid direct detection of bacterial toxins, CCD-based fluorescent activity analysis of microbial toxins and a simple ECL-based CCD detection system. However, while effective for molecular analysis, most of these technologies are not as sensitive as traditional culturing methods for detecting microorganisms. There is a need to combine optical biosensors with traditional methods to speed culture-based detection and to provide more information regarding the pathogens.

  6. Plasmonic biosensor for label-free G-quadruplexes detection

    NASA Astrophysics Data System (ADS)

    Qiu, Suyan; Zhao, Fusheng; Santos, Greggy M.; Shih, Wei-Chuan

    2016-03-01

    G-quadruplex, readily formed by the G-rich sequence, potentially distributes in over 40 % of all human genes, such as the telomeric DNA with the G-rich sequence found at the end of the chromosome. The G-quadruplex structure is supposed to possess a diverse set of critical functions in the mammalian genome for transcriptional regulation, DNA replication and genome stability. However, most of the currently available methods for G-quadruplex identification are restricted to fluorescence techniques susceptible to poor sensitivity. It is essential to propose methods with higher sensitivity to specifically recognize the G-quadruplexes. In this study, we demonstrate a label-free plasmonic biosensor for G-quadruplex detection by relying on the advantages of nanoporous gold (NPG) disks that provide high-density plasmonic hot spots, suitable for molecular recognition capability without the requirement for labeling processes.

  7. Orientational nanoparticle assemblies and biosensors.

    PubMed

    Ma, Wei; Xu, Liguang; Wang, Libing; Kuang, Hua; Xu, Chuanlai

    2016-05-15

    Assemblies of nanoparticles (NPs) have regional correlated properties with new features compared to individual NPs or random aggregates. The orientational NP assembly contributes greatly to the collective interaction of individual NPs with geometrical dependence. Therefore, orientational NPs assembly techniques have emerged as promising tools for controlling inorganic NPs spatial structures with enhanced interesting properties. The research fields of orientational NP assembly have developed rapidly with characteristics related to the different methods used, including chemical, physical and biological techniques. The current and potential applications, important challenges remain to be investigated. An overview of recent developments in orientational NPs assemblies, the multiple strategies, biosensors and challenges will be discussed in this review. PMID:26708241

  8. Microfabricated silicon biosensors for microphysiometry

    NASA Technical Reports Server (NTRS)

    Bousse, L. J.; Libby, J. M.; Parce, J. W.

    1993-01-01

    Microphysiometers are biosensor devices that measure the metabolic rate of living cells by detecting the rate of extracellular acidification caused by a small number of cells. The cells are entrapped in a microvolume chamber, whose bottom surface is a silicon sensor chip. In a further miniaturization step, we have recently fabricated multichannel flow-through chips that will allow greater throughput and multiplicity. Microphysiometer technology can be applied to the detection of microorganisms. We describe the sensitive detection of bacteria and yeast. Further applications of microphysiometry to the characterization of microorganisms can be anticipated.

  9. Built Environment Analysis Tool: April 2013

    SciTech Connect

    Porter, C.

    2013-05-01

    This documentation describes the tool development. It was created to evaluate the effects of built environment scenarios on transportation energy and greenhouse gas (GHG) emissions. This documentation also provides guidance on how to apply the tool.

  10. Principles of As-Built Engineering

    SciTech Connect

    Dolin, R.M.; Hefele, J.

    1996-11-01

    As-Built Engineering is a product realization methodology founded on the notion that life-cycle engineering should be based on what is actually produced and not on what is nominally designed. As-Built Engineering is a way of thinking about the production realization process that enables customization in mass production environments. It questions the relevance of nominal based methods of engineering and the role that tolerancing plays in product realization. As-Built Engineering recognizes that there will always be errors associated with manufacturing that cannot be controlled and therefore need to be captured in order to fully characterize each individual product`s unique attributes. One benefit of As-Built Engineering is the ability to provide actual product information to designers and analysts enabling them to verify their assumptions using actual part and assembly data. Another benefit is the ability to optimize new and re-engineered assemblies.

  11. The Built Environment Predicts Observed Physical Activity

    PubMed Central

    Kelly, Cheryl; Wilson, Jeffrey S.; Schootman, Mario; Clennin, Morgan; Baker, Elizabeth A.; Miller, Douglas K.

    2014-01-01

    Background: In order to improve our understanding of the relationship between the built environment and physical activity, it is important to identify associations between specific geographic characteristics and physical activity behaviors. Purpose: Examine relationships between observed physical activity behavior and measures of the built environment collected on 291 street segments in Indianapolis and St. Louis. Methods: Street segments were selected using a stratified geographic sampling design to ensure representation of neighborhoods with different land use and socioeconomic characteristics. Characteristics of the built environment on-street segments were audited using two methods: in-person field audits and audits based on interpretation of Google Street View imagery with each method blinded to results from the other. Segments were dichotomized as having a particular characteristic (e.g., sidewalk present or not) based on the two auditing methods separately. Counts of individuals engaged in different forms of physical activity on each segment were assessed using direct observation. Non-parametric statistics were used to compare counts of physically active individuals on each segment with built environment characteristic. Results: Counts of individuals engaged in physical activity were significantly higher on segments with mixed land use or all non-residential land use, and on segments with pedestrian infrastructure (e.g., crosswalks and sidewalks) and public transit. Conclusion: Several micro-level built environment characteristics were associated with physical activity. These data provide support for theories that suggest changing the built environment and related policies may encourage more physical activity. PMID:24904916

  12. Biosensor Approach to Psychopathology Classification

    PubMed Central

    Koshelev, Misha; Lohrenz, Terry; Vannucci, Marina; Montague, P. Read

    2010-01-01

    We used a multi-round, two-party exchange game in which a healthy subject played a subject diagnosed with a DSM-IV (Diagnostic and Statistics Manual-IV) disorder, and applied a Bayesian clustering approach to the behavior exhibited by the healthy subject. The goal was to characterize quantitatively the style of play elicited in the healthy subject (the proposer) by their DSM-diagnosed partner (the responder). The approach exploits the dynamics of the behavior elicited in the healthy proposer as a biosensor for cognitive features that characterize the psychopathology group at the other side of the interaction. Using a large cohort of subjects (n = 574), we found statistically significant clustering of proposers' behavior overlapping with a range of DSM-IV disorders including autism spectrum disorder, borderline personality disorder, attention deficit hyperactivity disorder, and major depressive disorder. To further validate these results, we developed a computer agent to replace the human subject in the proposer role (the biosensor) and show that it can also detect these same four DSM-defined disorders. These results suggest that the highly developed social sensitivities that humans bring to a two-party social exchange can be exploited and automated to detect important psychopathologies, using an interpersonal behavioral probe not directly related to the defining diagnostic criteria. PMID:20975934

  13. Comparative advantages of mechanical biosensors

    PubMed Central

    Arlett, J.L.; Myers, E.B.; Roukes, M.L.

    2013-01-01

    Mechanical interactions are fundamental to biology. Mechanical forces of chemical origin determine motility and adhesion on the cellular scale, and govern transport and affinity on the molecular scale. Biological sensing in the mechanical domain provides unique opportunities to measure forces, displacements and mass changes from cellular and subcellular processes. Nanomechanical systems are particularly well matched in size with molecular interactions, and provide a basis for biological probes with single-molecule sensitivity. Here we review micro- and nanoscale biosensors, with a particular focus on fast mechanical biosensing in fluid by mass- and force-based methods, and the challenges presented by non-specific interactions. We explain the general issues that will be critical to the success of any type of next-generation mechanical biosensor, such as the need to improve intrinsic device performance, fabrication reproducibility and system integration. We also discuss the need for a greater understanding of analyte–sensor interactions on the nanoscale and of stochastic processes in the sensing environment. PMID:21441911

  14. Early Lung Cancer Diagnosis by Biosensors

    PubMed Central

    Zhang, Yuqian; Yang, Dongliang; Weng, Lixing; Wang, Lianhui

    2013-01-01

    Lung cancer causes an extreme threat to human health, and the mortality rate due to lung cancer has not decreased during the last decade. Prognosis or early diagnosis could help reduce the mortality rate. If microRNA and tumor-associated antigens (TAAs), as well as the corresponding autoantibodies, can be detected prior to clinical diagnosis, such high sensitivity of biosensors makes the early diagnosis and prognosis of cancer realizable. This review provides an overview of tumor-associated biomarker identifying methods and the biosensor technology available today. Laboratorial researches utilizing biosensors for early lung cancer diagnosis will be highlighted. PMID:23892596

  15. Remote sensing using an airborne biosensor

    SciTech Connect

    Ligler, F.S.; Anderson, G.P.; Davidson, P.T.; Stenger, D.A.; Ives, J.T.; King, K.D.; Page, G.; Whelan, J.P.

    1998-08-15

    There is no current method for remote identification of aerosolized bacteria. In particular, such a capability is required to warn of a biological warfare attack prior to human exposure. A fiber optic biosensor, capable of running four simultaneous immunoassays, was integrated with an automated fluidics unit, a cyclone-type air sampler, a radio transceiver, and batteries on a small, remotely piloted airplane capable of carrying a 4.5-kg payload. The biosensor system was able to collect aerosolized bacteria in flight, identify them, and transmit the data to the operator on the ground. The results demonstrate the feasibility of integrating a biosensor into a portable, remotely operated system for environmental analysis.

  16. 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.

  17. Determination of endotoxin through an aptamer-based impedance biosensor.

    PubMed

    Su, Wenqiong; Lin, Meng; Lee, Hyuck; Cho, MiSuk; Choe, Woo-Seok; Lee, Youngkwan

    2012-02-15

    Lipopolysaccharide (LPS) often referred to endotoxin is an undesirable impurity frequently entrained with various recombinant protein therapeutics and plasmid DNA (pDNA) vaccines of bacterial origin. The inherent toxicities (e.g. fever, hypotension, shock and death) of LPS render its early and sensitive detection essential for several biological assays and/or parenteral administrations of biotherapeutics. In this study, an electrochemical biosensor using an LPS specific single stranded DNA (ssDNA) aptamer as a probe was developed. Amine-terminated aptamer exhibiting high affinity (K(d)=11.9 nM) to LPS was immobilized on a gold electrode using 3-mercaptopropionic acid (MPA) as a linker. Each step of the modification process was characterized by cyclic voltammetry (CV) and electrochemical impendence spectroscopy (EIS). A good linear relationship of the changes in the charge-transfer resistance (ΔR(et)) and the logarithmic value of LPS concentration was demonstrated in a broad dynamic detection range of 0.001-1 ng/ml. Furthermore, the aptasensor showed a high selectivity to LPS despite the presence of pDNA, RNA and bovine serum albumin (BSA) and could be regenerated in low pH condition, offering a promising option for detecting LPS often present in a complex milieu. PMID:22182428

  18. 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

  19. Quantitative Investigation of Protein-Nucleic Acid Interactions by Biosensor Surface Plasmon Resonance

    PubMed Central

    Wang, Shuo; Poon, Gregory M. K.; Wilson, W. David

    2015-01-01

    Biosensor-surface plasmon resonance (SPR) technology has emerged as a powerful label-free approach for the study of nucleic acid interactions in real time. The method provides simultaneous equilibrium and kinetic characterization for biomolecular interactions with minimal materials and without an external probe. A detailed and practical guide for protein-DNA interaction analyses using biosensor-SPR methods is presented. Details of the SPR technology and basic fundamentals are described with recommendations on the preparation of the SPR instrument, sensor chips and samples, as well as extensive information on experimental design, quantitative and qualitative data analyses and presentation. A specific example of the interaction of a transcription factor with DNA is shown with results evaluated by both kinetic and steady-state SPR methods. PMID:26404159

  20. Electronic transport in methylated fragments of DNA

    NASA Astrophysics Data System (ADS)

    de Almeida, M. L.; Oliveira, J. I. N.; Lima Neto, J. X.; Gomes, C. E. M.; Fulco, U. L.; Albuquerque, E. L.; Freire, V. N.; Caetano, E. W. S.; de Moura, F. A. B. F.; Lyra, M. L.

    2015-11-01

    We investigate the electronic transport properties of methylated deoxyribonucleic-acid (DNA) strands, a biological system in which methyl groups are added to DNA (a major epigenetic modification in gene expression), sandwiched between two metallic platinum electrodes. Our theoretical simulations apply an effective Hamiltonian based on a tight-binding model to obtain current-voltage curves related to the non-methylated/methylated DNA strands. The results suggest potential applications in the development of novel biosensors for molecular diagnostics.

  1. Electronic transport in methylated fragments of DNA

    SciTech Connect

    Almeida, M. L. de; Oliveira, J. I. N.; Lima Neto, J. X.; Gomes, C. E. M.; Fulco, U. L. Albuquerque, E. L.; Freire, V. N.; Caetano, E. W. S.; Moura, F. A. B. F. de; Lyra, M. L.

    2015-11-16

    We investigate the electronic transport properties of methylated deoxyribonucleic-acid (DNA) strands, a biological system in which methyl groups are added to DNA (a major epigenetic modification in gene expression), sandwiched between two metallic platinum electrodes. Our theoretical simulations apply an effective Hamiltonian based on a tight-binding model to obtain current-voltage curves related to the non-methylated/methylated DNA strands. The results suggest potential applications in the development of novel biosensors for molecular diagnostics.

  2. Recent Advances in Nanotechnology Applied to Biosensors

    PubMed Central

    Zhang, Xueqing; Guo, Qin; Cui, Daxiang

    2009-01-01

    In recent years there has been great progress the application of nanomaterials in biosensors. The importance of these to the fundamental development of biosensors has been recognized. In particular, nanomaterials such as gold nanoparticles, carbon nanotubes, magnetic nanoparticles and quantum dots have been being actively investigated for their applications in biosensors, which have become a new interdisciplinary frontier between biological detection and material science. Here we review some of the main advances in this field over the past few years, explore the application prospects, and discuss the issues, approaches, and challenges, with the aim of stimulating a broader interest in developing nanomaterial-based biosensors and improving their applications in disease diagnosis and food safety examination. PMID:22399954

  3. PRINCIPLES OF AFFINITY-BASED BIOSENSORS

    EPA Science Inventory

    Despite the amount of resources that have been invested by national and international academic, government, and commercial sectors to develop affinity-based biosensor products, little obvious success has been realized through commercialization of these devices for specific applic...

  4. Non-antibody protein-based biosensors.

    PubMed

    Ko Ferrigno, Paul

    2016-06-30

    Biosensors that depend on a physical or chemical measurement can be adversely affected by non-specific interactions. For example, a biosensor designed to measure specifically the levels of a rare analyte can give false positive results if there is even a small amount of interaction with a highly abundant but irrelevant molecule. To overcome this limitation, the biosensor community has frequently turned to antibody molecules as recognition elements because they are renowned for their exquisite specificity. Unfortunately antibodies can often fail when immobilised on inorganic surfaces, and alternative biological recognition elements are needed. This article reviews the available non-antibody-binding proteins that have been successfully used in electrical and micro-mechanical biosensor platforms. PMID:27365032

  5. Non-antibody protein-based biosensors

    PubMed Central

    2016-01-01

    Biosensors that depend on a physical or chemical measurement can be adversely affected by non-specific interactions. For example, a biosensor designed to measure specifically the levels of a rare analyte can give false positive results if there is even a small amount of interaction with a highly abundant but irrelevant molecule. To overcome this limitation, the biosensor community has frequently turned to antibody molecules as recognition elements because they are renowned for their exquisite specificity. Unfortunately antibodies can often fail when immobilised on inorganic surfaces, and alternative biological recognition elements are needed. This article reviews the available non-antibody-binding proteins that have been successfully used in electrical and micro-mechanical biosensor platforms. PMID:27365032

  6. Enzyme Nanoparticles-Based Electronic Biosensor

    SciTech Connect

    Liu, Guodong; Lin, Yuehe; Ostatna, V.; Wang, Joseph

    2005-06-28

    A novel method for fabricating electronic biosensors based on coupling enzyme nanoparticles and self assembly technology is illustrated. Redox horseradish peroxidase nanoparticles were prepared by desolvation with ethanol and subsequent crosslinking with glutaraldehyde. The cross-linked enzyme nanoparticles were functionalized by cysteine to introduce thiol groups on the nanoparticle surface. Immobilized enzyme nanoparticle on the gold electrode by self-assembly kept redox and electrocatalytic activities, and was used to develop reagentless biosensors for H2O2 detection without promoters and mediators. The new approach is simple, low cost and circumvents complications associated with solution systems. It is a universal immobilization method for biosensor, biomedical devices, biofuel cells and enzymatic bioreactors fabrication and expected to open new opportunities for biosensor, clinical diagnostics, and for bioanalysis, in general.

  7. Biosensors for Inorganic and Organic Arsenicals

    PubMed Central

    Chen, Jian; Rosen, Barry P.

    2014-01-01

    Arsenic is a natural environmental contaminant to which humans are routinely exposed and is strongly associated with human health problems, including cancer, cardiovascular and neurological diseases. To date, a number of biosensors for the detection of arsenic involving the coupling of biological engineering and electrochemical techniques has been developed. The properties of whole-cell bacterial or cell-free biosensors are summarized in the present review with emphasis on their sensitivity and selectivity. Their limitations and future challenges are highlighted. PMID:25587436

  8. 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.

  9. Built Environment Energy Analysis Tool Overview (Presentation)

    SciTech Connect

    Porter, C.

    2013-04-01

    This presentation provides an overview of the Built Environment Energy Analysis Tool, which is designed to assess impacts of future land use/built environment patterns on transportation-related energy use and greenhouse gas (GHG) emissions. The tool can be used to evaluate a range of population distribution and urban design scenarios for 2030 and 2050. This tool was produced as part of the Transportation Energy Futures (TEF) project, a Department of Energy-sponsored multi-agency project initiated to pinpoint underexplored strategies for abating GHGs and reducing petroleum dependence related to transportation.

  10. Integrated optical biosensor system (IOBS)

    DOEpatents

    Grace, Karen M.; Sweet, Martin R.; Goeller, Roy M.; Morrison, Leland Jean; Grace, Wynne Kevin; Kolar, Jerome D.

    2007-10-30

    An optical biosensor has a first enclosure with a pathogen recognition surface, including a planar optical waveguide and grating located in the first enclosure. An aperture is in the first enclosure for insertion of sample to be investigated to a position in close proximity to the pathogen recognition surface. A laser in the first enclosure includes means for aligning and means for modulating the laser, the laser having its light output directed toward said grating. Detection means are located in the first enclosure and in optical communication with the pathogen recognition surface for detecting pathogens after interrogation by the laser light and outputting the detection. Electronic means is located in the first enclosure and receives the detection for processing the detection and outputting information on the detection, and an electrical power supply is located in the first enclosure for supplying power to the laser, the detection means and the electronic means.

  11. Locked nucleic acid based beacons for surface interaction studies and biosensor development

    PubMed Central

    Martinez, Karen; Estevez, M.-Carmen; Wu, Yanrong; Phillips, Joseph A.; Medley, Colin D.; Tan, Weihong

    2011-01-01

    DNA sensors and microarrays permit fast, simple and real-time detection of nucleic acids through the design and use of increasingly sensitive, selective and robust molecular probes. Specifically, molecular beacons (MBs) have been employed for this purpose; however, their potential in the development of solid-surface-based biosensors has not been fully realized. This is mainly a consequence of the beacon’s poor stability due to the hairpin structure once immobilized onto a solid surface, commonly resulting in a low signal enhancement. Here, we report the design of a new MB that overcomes some of the limitations of MBs for surface immobilization. Essentially, this new design adds locked nucleic acid bases (LNAs) to the beacon structure, resulting in a LNA molecular beacon (LMB) with robust stability after surface immobilization. To test the efficacy of LMBs against that of regular molecular beacons (RMBs), the properties of selectivity, sensitivity, thermal stability, hybridization kinetics and robustness for the detection of target sequences were compared and evaluated. A 25-fold enhancement was achieved for the LMB on surface with detection limits reaching the low nanomolar range. In addition, the LMB-based biosensor was shown to possess better stability, reproducibility, selectivity and robustness when compared to the RMB. Therefore, as an alternative to conventional DNA and as a prospective tool for use in both DNA microarrays and biosensors, these results demonstrate the potential of the locked nucleic acid bases for nucleic acid design for surface immobilization. PMID:19351140

  12. Angle-resolved diffraction grating biosensor based on porous silicon

    NASA Astrophysics Data System (ADS)

    Lv, Changwu; Jia, Zhenhong; Liu, Yajun; Mo, Jiaqing; Li, Peng; Lv, Xiaoyi

    2016-03-01

    In this study, an optical biosensor based on a porous silicon composite structure was fabricated using a simple method. This structure consists of a thin, porous silicon surface diffraction grating and a one-dimensional porous silicon photonic crystal. An angle-resolved diffraction efficiency spectrum was obtained by measuring the diffraction efficiency at a range of incident angles. The angle-resolved diffraction efficiency of the 2nd and 3rd orders was studied experimentally and theoretically. The device was sensitive to the change of refractive index in the presence of a biomolecule indicated by the shift of the diffraction efficiency spectrum. The sensitivity of this sensor was investigated through use of an 8 base pair antifreeze protein DNA hybridization. The shifts of the angle-resolved diffraction efficiency spectrum showed a relationship with the change of the refractive index, and the detection limit of the biosensor reached 41.7 nM. This optical device is highly sensitive, inexpensive, and simple to fabricate. Using shifts in diffraction efficiency spectrum to detect biological molecules has not yet been explored, so this study establishes a foundation for future work.

  13. ADMET biosensors: up-to-date issues and strategies.

    PubMed

    Fang, Yan; Offenhaeusser, Andrease

    2004-12-01

    This insight review introduces the new concepts, theories, technology, instruments, frontier issues, and key strategies of ADMET (absorption, distribution, metabolism, elimination, and toxicity) biosensors, from the fermi to the quantum levels. Information about ADMET, originating from one author's invention, a patented pharmacotherapy for rescuing cardio-cerebral vascular stunning and regulating vascular endothelial growth-factor signaling at the post-genomic level, can be detected by a new generation of ADMET biosensor. This is a single-cell/single-molecule field-effect transistor (FET) hybrid system, where single molecules or single cells are assembled at the FET surface in a high density array manner via complementary metal-oxide-semiconductor (CMOS)-compatible technologies. Within a given nanometer distance, ADMET-mediated oxidation-reduction (redox) potentials, electrochemistry responses, and electron transfer processes can be simultaneously and directly probed by the gates of field-effect transistor arrays. The nanometer details of the functional coupling principles and characterization technologies of DNA single-molecule/single-cell FETs, as well as the design of lab-on-a-chip instruments, are indicated. Four frontier issues and key strategies are elucidated in detail. This can lead to innovative technology for high-throughout screening of labs-on-chips to resolve the pharmaceutical industry's current bottleneck via novel, FET-based drug discovery and single-molecule/single-cell screening methods, which can bring about a pharmaceutical industry revolution in the 21st century. PMID:15567991

  14. Integration of biosensors into digital microfluidics: Impact of hydrophilic surface of biosensors on droplet manipulation.

    PubMed

    Samiei, Ehsan; Luka, George S; Najjaran, Homayoun; Hoorfar, Mina

    2016-07-15

    Several studies have been performed on the integration of biosensors into digital microfluidics (DMF). Despite the general success in their detection capabilities, there are still two challenges associated with the integration of biosensors into DMF: (1) complete removal of the droplet containing the analytes from the sensing surface; and (2) biochemical regeneration of the biosensor involving detaching the target analyte from the receptor after each round of sensing. The latter is case dependent and the solution can vary from one application to another. Our research aims at addressing the former, the solution to which is applicable to all biosensors integrated to DMF. This paper presents a thorough characterization of the hydrophilic surface of the biosensor in terms of wettability and geometry, taking into account the overall configuration of the DMF platform. Consequently, we identify the optimal geometry of the sensing surface and the DMF platform providing successful removal of the target droplet from the sensing surface after detection. Based on the results, the gap height is suggested to be chosen at the upper limit of the applicable range. Also, the biosensor, patterned on the device top plate, is recommended to be designed with a high aspect ratio and aligned with the center of the actuating electrode. As a proof of concept, the optimum configuration is implemented on a DMF platform with an interdigitated capacitive biosensor to detect different concentrations of Cryptosporidium, for which it is shown that the sample droplet is removed successfully from the superhydrophilic surface of the biosensor. PMID:27016626

  15. "Anarchy and a Built-In Gyro"

    ERIC Educational Resources Information Center

    Healy, Timothy S.

    1978-01-01

    Any good college resists rule, but it also has a built-in self-correcting apparatus, suggests this university president. Trustees should ask themselves: Does your college surprise you? Do you trust it? Do you wish you could be more a part of it? (Author/LBH)

  16. Multichannel Analyzer Built from a Microcomputer.

    ERIC Educational Resources Information Center

    Spencer, C. D.; Mueller, P.

    1979-01-01

    Describes a multichannel analyzer built using eight-bit S-100 bus microcomputer hardware. The output modes are an oscilloscope display, print data, and send data to another computer. Discusses the system's hardware, software, costs, and advantages relative to commercial multichannels. (Author/GA)

  17. Schooling Built on the Multiple Intelligences

    ERIC Educational Resources Information Center

    Kunkel, Christine D.

    2009-01-01

    This article features a school built on multiple intelligences. As the first multiple intelligences school in the world, the Key Learning Community shapes its students' days to include significant time in the musical, spatial and bodily-kinesthetic intelligences, as well as the more traditional areas of logical-mathematical and linguistics. In…

  18. 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

  19. Development of an acoustic wave based biosensor for vapor phase detection of small molecules

    NASA Astrophysics Data System (ADS)

    Stubbs, Desmond

    For centuries scientific ingenuity and innovation have been influenced by Mother Nature's perfect design. One of her more elusive designs is that of the sensory olfactory system, an array of highly sensitive receptors responsible for chemical vapor recognition. In the animal kingdom this ability is magnified among canines where ppt (parts per trillion) sensitivity values have been reported. Today, detection dogs are considered an essential part of the US drug and explosives detection schemes. However, growing concerns about their susceptibility to extraneous odors have inspired the development of highly sensitive analytical detection tools or biosensors known as "electronic noses". In general, biosensors are distinguished from chemical sensors in that they use an entity of biological origin (e.g. antibody, cell, enzyme) immobilized onto a surface as the chemically-sensitive film on the device. The colloquial view is that the term "biosensors" refers to devices which detect the presence of entities of biological origin, such as proteins or single-stranded DNA and that this detection must take place in a liquid. Our biosensor utilizes biomolecules, specifically IgG monoclonal antibodies, to achieve molecular recognition of relatively small molecules in the vapor phase.

  20. Amperometric Glucose Biosensor Based on Self-Assembling Glucose Oxidase on Carbon Nanotubes

    SciTech Connect

    Liu, Guodong; Lin, Yuehe

    2006-01-01

    A flow injection amperometric glucose biosensor based on electrostatic self-assembling glucose oxidase (GOx) on a carbon nanotube (CNT)-modified glassy carbon transducer is described. GOx is immobilized on the negatively charged CNT surface by alternatively assembling a cationic polydiallyldimethylammonium chloride (PDDA) layer and a GOx layer. The unique sandwich-like layer structure (PDDA/GOx/PDDA/CNT) formed by self-assembling provides a favorable microenvironment to keep the bioactivity of GOx and to prevent enzyme molecule leakage. The direct electrochemistry behavior of GOx and electrocatalysis of H2O2 on the fabricated PDDA/GOx/PDDA/CNT electrode demonstrated that such a biosensor fabrication method preserves the activity of enzyme molecules and the mechanical and electrocatalytic properties of carbon nanotubes, enabling sensitive determination of glucose. Flow injection amperometric detection of glucose is carried out at -100 mV (vs Ag/AgCl) in 0.05 M phosphate buffer solution (pH 7.4) with wide linear response range of 15 uM- 6 mM and a detection limit of 7 uM. The PDDA/GOx/PDDA/CNT/GC biosensor showed excellent properties for the sensitive determination of glucose with good reproducibility, remarkable stability, and free of interference from other co-existing electroactive species. The present methods can be applied to assemble other enzyme molecules and biological molecules, such as antibody, antigen, and DNA, to the CNT surface for wide biosensor and bioassay applications.

  1. FRET based biosensor for detection of active NF-kB

    NASA Astrophysics Data System (ADS)

    Baldini, Francesco; Citti, Lorenzo; Domenici, Claudio; Giannetti, Ambra; Tedeschi, Lorena; Vo-Dinh, Tuan; Wabuyele, Musundi B.

    2005-05-01

    The Nuclear Factor kB is a transcription factor, ubiquitously expressed, involved in the regulation of a large number of genes and in a variety of human disease including inflammation, asthma, atherosclerosis, AIDS, septic shock, arthritis and cancer. The critical need for a simple and direct method to evaluate the quantity of active NF-kB in a biological sample can be addressed using a suitable and reusable biosensor. For this purpose, a novel method, using fluorescence resonance energy transfer (FRET), to detect the active form of NF-kB binding a specific DNA sequence has been developed. A single-stranded DNA (ssDNA) with auto-complementary sequence has been properly designed and synthesized. In order to evaluate FRET due to the DNA/protein binding interaction taking place between double-stranded DNA (dsDNA) immobilized in a capillary wall and NF-kB proteins, a highly sensitive FRET-based biosensor system developed in our laboratory was used. Preliminary results show that our system was capable of detecting the active form of NF-kB protein with a detection efficiency of about 90% and that the system has a good regenerability.

  2. SIRE-technology-based biosensors: will they do the job?

    NASA Astrophysics Data System (ADS)

    Kriz, Dario

    1997-06-01

    A new biosensor technology (SIRE--sensors based on injectable recognition elements) is described. Its application in laboratory equipment, medical survey equipment and process monitoring is reviewed. Furthermore, the promising practical and commercial relevance of SIRE- Biosensors is discussed.

  3. Optimization of Xenon Biosensors for Detection of ProteinInteractions

    SciTech Connect

    Lowery, Thomas J.; Garcia, Sandra; Chavez, Lana; Ruiz, E.Janette; Wu, Tom; Brotin, Thierry; Dutasta, Jean-Pierre; King, David S.; Schultz, Peter G.; Pines, Alex; Wemmer, David E..

    2005-08-03

    Hyperpolarized 129Xe NMR can detect the presence of specific low-concentration biomolecular analytes by means of the xenon biosensor, which consists of a water-soluble, targeted cryptophane-A cage that encapsulates xenon. In this work we use the prototypical biotinylated xenon biosensor to determine the relationship between the molecular composition of the xenon biosensor and the characteristics of protein-bound resonances. The effects of diastereomer overlap, dipole-dipole coupling, chemical shift anisotropy, xenon exchange, and biosensor conformational exchange on protein-bound biosensor signal were assessed. It was found that optimal protein-bound biosensor signal can be obtained by minimizing the number of biosensor diastereomers and using a flexible linker of appropriate length. Both the linewidth and sensitivity of chemical shift to protein binding of the xenon biosensor were found to be inversely proportional to linker length.

  4. Recovery Based Nanowire Field-Effect Transistor Detection of Pathogenic Avian Influenza DNA

    NASA Astrophysics Data System (ADS)

    Lin, Chih-Heng; Chu, Chia-Jung; Teng, Kang-Ning; Su, Yi-Jr; Chen, Chii-Dong; Tsai, Li-Chu; Yang, Yuh-Shyong

    2012-02-01

    Fast and accurate diagnosis is critical in infectious disease surveillance and management. We proposed a DNA recovery system that can easily be adapted to DNA chip or DNA biosensor for fast identification and confirmation of target DNA. This method was based on the re-hybridization of DNA target with a recovery DNA to free the DNA probe. Functionalized silicon nanowire field-effect transistor (SiNW FET) was demonstrated to monitor such specific DNA-DNA interaction using high pathogenic strain virus hemagglutinin 1 (H1) DNA of avian influenza (AI) as target. Specific electric changes were observed in real-time for AI virus DNA sensing and device recovery when nanowire surface of SiNW FET was modified with complementary captured DNA probe. The recovery based SiNW FET biosensor can be further developed for fast identification and further confirmation of a variety of influenza virus strains and other infectious diseases.

  5. 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

  6. Low cost flatbed scanner label-free biosensor

    NASA Astrophysics Data System (ADS)

    Aygun, Ugur; Avci, Oguzhan; Seymour, Elif; Sevenler, Derin D.; Urey, Hakan; Ünlü, M. Selim; Ozkumur, Ayca Yalcin

    2016-03-01

    In this paper, we demonstrate utilization of a commercial flatbed document scanner as a label-free biosensor for highthroughput imaging of DNA and protein microarrays. We implemented an interferometric sensing technique through use of a silicon/oxide layered substrate, and easy to implement hardware modifications such as re-aligning moving parts and inserting a custom made sample plate. With a cost as low as 100USD, powered by a USB cable, and scan speed of 30 seconds for a 4mm x 4 mm area with ~10μm lateral resolution, the presented system offers a super low cost, easy to use alternative to commercially available label-free systems.

  7. Measuring the Built Environment for Physical Activity

    PubMed Central

    Brownson, Ross C.; Hoehner, Christine M.; Day, Kristen; Forsyth, Ann; Sallis, James F.

    2009-01-01

    Physical inactivity is one of the most important public health issues in the U.S. and internationally. Increasingly, links are being identified between various elements of the physical—or built—environment and physical activity. To understand the impact of the built environment on physical activity, the development of high-quality measures is essential. Three categories of built environment data are being used: (1) perceived measures obtained by telephone interview or self-administered questionnaires; (2) observational measures obtained using systematic observational methods (audits); and (3) archival data sets that are often layered and analyzed with GIS. This review provides a critical assessment of these three types of built-environment measures relevant to the study of physical activity. Among perceived measures, 19 questionnaires were reviewed, ranging in length from 7 to 68 questions. Twenty audit tools were reviewed that cover community environments (i.e., neighborhoods, cities), parks, and trails. For GIS-derived measures, more than 50 studies were reviewed. A large degree of variability was found in the operationalization of common GIS measures, which include population density, land-use mix, access to recreational facilities, and street pattern. This first comprehensive examination of built-environment measures demonstrates considerable progress over the past decade, showing diverse environmental variables available that use multiple modes of assessment. Most can be considered first-generation measures, so further development is needed. In particular, further research is needed to improve the technical quality of measures, understand the relevance to various population groups, and understand the utility of measures for science and public health. PMID:19285216

  8. Device considerations for development of conductance-based biosensors

    PubMed Central

    Lee, Kangho; Nair, Pradeep R.; Scott, Adina; Alam, Muhammad A.; Janes, David B.

    2009-01-01

    Design and fabrication of electronic biosensors based on field-effect-transistor (FET) devices require understanding of interactions between semiconductor surfaces and organic biomolecules. From this perspective, we review practical considerations for electronic biosensors with emphasis on molecular passivation effects on FET device characteristics upon immobilization of organic molecules and an electrostatic model for FET-based biosensors. PMID:24753627

  9. Built-Environment Wind Turbine Roadmap

    SciTech Connect

    Smith, J.; Forsyth, T.; Sinclair, K.; Oteri, F.

    2012-11-01

    Although only a small contributor to total electricity production needs, built-environment wind turbines (BWTs) nonetheless have the potential to influence the public's consideration of renewable energy, and wind energy in particular. Higher population concentrations in urban environments offer greater opportunities for project visibility and an opportunity to acquaint large numbers of people to the advantages of wind projects on a larger scale. However, turbine failures will be equally visible and could have a negative effect on public perception of wind technology. This roadmap provides a framework for achieving the vision set forth by the attendees of the Built-Environment Wind Turbine Workshop on August 11 - 12, 2010, at the U.S. Department of Energy's National Renewable Energy Laboratory. The BWT roadmap outlines the stakeholder actions that could be taken to overcome the barriers identified. The actions are categorized as near-term (0 - 3 years), medium-term (4 - 7 years), and both near- and medium-term (requiring immediate to medium-term effort). To accomplish these actions, a strategic approach was developed that identifies two focus areas: understanding the built-environment wind resource and developing testing and design standards. The authors summarize the expertise and resources required in these areas.

  10. Contributions of built environment to childhood obesity.

    PubMed

    Rahman, Tamanna; Cushing, Rachel A; Jackson, Richard J

    2011-01-01

    As childhood obesity has reached epidemic proportions, it is critical to devise interventions that target the root causes of obesity and its risk factors. The two main components of childhood obesity are physical inactivity and improper nutrition, and it is becoming increasingly evident that the built environment can determine the level of exposure to these risk factors. Through a multidisciplinary literature review, we investigated the association between various built environment attributes and childhood obesity. We found that neighborhood features such as walkability/bikeability, mixed land use, accessible destinations, and transit increase resident physical activity; also that access to high-caloric foods and convenience stores increases risk of overweight and obesity, whereas the presence of neighborhood supermarkets and farmers' markets is associated with lower childhood body mass index and overweight status. It is evident that a child's built environment impacts his access to nutritious foods and physical activity. In order for children, as well as adults, to prevent onset of overweight or obesity, they need safe places to be active and local markets that offer affordable, healthy food options. Interventions that are designed to provide safe, walkable neighborhoods with access to necessary destinations will be effective in combating the epidemic of obesity. PMID:21259262

  11. Mobility disability and the urban built environment.

    PubMed

    Clarke, Philippa; Ailshire, Jennifer A; Bader, Michael; Morenoff, Jeffrey D; House, James S

    2008-09-01

    Research on the effects of the built environment in the pathway from impairment to disability has been largely absent. Using data from the Chicago Community Adult Health Study (2001-2003), the authors examined the effect of built environment characteristics on mobility disability among adults aged 45 or more years (n = 1,195) according to their level of lower extremity physical impairment. Built environment characteristics were assessed by using systematic social observation to independently rate street and sidewalk quality in the block surrounding each respondent's residence in the city of Chicago (Illinois). Using multinomial logistic regression, the authors found that street conditions had no effect on outdoor mobility among adults with only mild or no physical impairment. However, among adults with more severe impairment in neuromuscular and movement-related functions, the difference in the odd ratios for reporting severe mobility disability was over four times greater when at least one street was in fair or poor condition (characterized by cracks, potholes, or broken curbs). When all streets were in good condition, the odds of reporting mobility disability were attenuated in those with lower extremity impairment. If street quality could be improved, even somewhat, for those adults at greatest risk for disability in outdoor mobility, the disablement process could be slowed or even reversed. PMID:18667526

  12. Natural bacterial communities serve as quantitative geochemical biosensors

    DOE PAGESBeta

    Smith, Mark B.; Rocha, Andrea M.; Smillie, Chris S.; Olesen, Scott W.; Paradis, Charles; Wu, Liyou; Campbell, James H.; Fortney, Julian L.; Mehlhorn, Tonia L.; Lowe, Kenneth A.; et al

    2015-05-12

    Biological sensors can be engineered to measure a wide range of environmental conditions. Here we show that statistical analysis of DNA from natural microbial communities can be used to accurately identify environmental contaminants, including uranium and nitrate at a nuclear waste site. In addition to contamination, sequence data from the 16S rRNA gene alone can quantitatively predict a rich catalogue of 26 geochemical features collected from 93 wells with highly differing geochemistry characteristics. We extend this approach to identify sites contaminated with hydrocarbons from the Deepwater Horizon oil spill, finding that altered bacterial communities encode a memory of prior contamination,more » even after the contaminants themselves have been fully degraded. We show that the bacterial strains that are most useful for detecting oil and uranium are known to interact with these substrates, indicating that this statistical approach uncovers ecologically meaningful interactions consistent with previous experimental observations. Future efforts should focus on evaluating the geographical generalizability of these associations. Taken as a whole, these results indicate that ubiquitous, natural bacterial communities can be used as in situ environmental sensors that respond to and capture perturbations caused by human impacts. These in situ biosensors rely on environmental selection rather than directed engineering, and so this approach could be rapidly deployed and scaled as sequencing technology continues to become faster, simpler, and less expensive. Here we show that DNA from natural bacterial communities can be used as a quantitative biosensor to accurately distinguish unpolluted sites from those contaminated with uranium, nitrate, or oil. These results indicate that bacterial communities can be used as environmental sensors that respond to and capture perturbations caused by human impacts.« less

  13. Natural bacterial communities serve as quantitative geochemical biosensors

    SciTech Connect

    Smith, Mark B.; Rocha, Andrea M.; Smillie, Chris S.; Olesen, Scott W.; Paradis, Charles; Wu, Liyou; Campbell, James H.; Fortney, Julian L.; Mehlhorn, Tonia L.; Lowe, Kenneth A.; Earles, Jennifer E.; Phillips, Jana; Techtmann, Steve M.; Joyner, Dominique C.; Elias, Dwayne A.; Bailey, Kathryn L.; Hurt, Richard A.; Preheim, Sarah P.; Sanders, Matthew C.; Yang, Joy; Mueller, Marcella A.; Brooks, Scott; Watson, David B.; Zhang, Ping; He, Zhili; Dubinsky, Eric A.; Adams, Paul D.; Arkin, Adam P.; Fields, Matthew W.; Zhou, Jizhong; Alm, Eric J.; Hazen, Terry C.

    2015-05-12

    Biological sensors can be engineered to measure a wide range of environmental conditions. Here we show that statistical analysis of DNA from natural microbial communities can be used to accurately identify environmental contaminants, including uranium and nitrate at a nuclear waste site. In addition to contamination, sequence data from the 16S rRNA gene alone can quantitatively predict a rich catalogue of 26 geochemical features collected from 93 wells with highly differing geochemistry characteristics. We extend this approach to identify sites contaminated with hydrocarbons from the Deepwater Horizon oil spill, finding that altered bacterial communities encode a memory of prior contamination, even after the contaminants themselves have been fully degraded. We show that the bacterial strains that are most useful for detecting oil and uranium are known to interact with these substrates, indicating that this statistical approach uncovers ecologically meaningful interactions consistent with previous experimental observations. Future efforts should focus on evaluating the geographical generalizability of these associations. Taken as a whole, these results indicate that ubiquitous, natural bacterial communities can be used as in situ environmental sensors that respond to and capture perturbations caused by human impacts. These in situ biosensors rely on environmental selection rather than directed engineering, and so this approach could be rapidly deployed and scaled as sequencing technology continues to become faster, simpler, and less expensive. Here we show that DNA from natural bacterial communities can be used as a quantitative biosensor to accurately distinguish unpolluted sites from those contaminated with uranium, nitrate, or oil. These results indicate that bacterial communities can be used as environmental sensors that respond to and capture perturbations caused by human impacts.

  14. Single-molecule surface studies of fibrinogen and DNA on semiconductors

    NASA Astrophysics Data System (ADS)

    Kong, Xianhua

    Understanding of protein adsorption onto non-biological substrates is of fundamental interest in science, but also has great potential technological applications in medical devices and biosensors. This study explores the non-specific interaction, at the single molecule level, of a blood protein and DNA with semiconductor surfaces through the use of a custom built, non rastering electron emission microscope and a scanning probe microscope. The specifics and history of electron emission are described as well as the equipment used in this study. The protein examined in this study is human plasma fibrinogen, which plays an important role in haemostatis and thrombosis, and deoxyribonucleic acid (DNA) is also studied. A novel technique for determining the photothreshold of biomolecules on single molecule level is developed and applied to fibrinogen molecules adsorbed on oxidized silicon surfaces, using photo-electron emission microscopy (PEEM). Three theoretical models are employed and compared to analyze the experimental photothreshold data. The non-specific adsorption of human plasma fibrinogen on oxidized p- and n- type silicon (100) surfaces is investigated to characterize both hydrophobic interactions and electrostatic forces. The experimental results indicate that hydrophobic interactions are one of the driving forces for protein adsorption and the electrostatic interactions also play a role in the height of the fibrinogen molecules adsorbed on the surface. PEEM images establish a photo threshold of 5.0 +/- 0.2 eV for fibrinogen on both n-type and p-type Si (100) surfaces. We suggest that the photothreshold results from surface state associated Fermi level (EF) pinning and there exists negative charge transfer from the adsorbed fibrinogen onto the p-type silicon substrates, while on n-type silicon substrates negative charge is transferred in the opposite direction. The adsorption of deoxyribonucleic acid (DNA) on mica and silicon is studied in liquid and ambient

  15. 2. EAST ELEVATION OF IPA FACTORY; TWOSTORY SECTION BUILT IN ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    2. EAST ELEVATION OF IPA FACTORY; TWO-STORY SECTION BUILT IN 1892 AND PARTIALLY DESTROYED PARAPET SECTION BUILT CA. 1948. BRICK CHIMNEY ALSO BUILT CA. 1948. - Illinois Pure Aluminum Company, 109 Holmes Street, Lemont, Cook County, IL

  16. One Bedroom Units: Floor Plan, South Elevation (As Built), North ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    One Bedroom Units: Floor Plan, South Elevation (As Built), North Elevation (As Built), Section A-A (As Built), Section AA (Existing) - Aluminum City Terrace, East Hill Drive, New Kensington, Westmoreland County, PA

  17. Nanomaterial-mediated Biosensors for Monitoring Glucose

    PubMed Central

    Taguchi, Masashige; Ptitsyn, Andre; McLamore, Eric S.

    2014-01-01

    Real-time monitoring of physiological glucose transport is crucial for gaining new understanding of diabetes. Many techniques and equipment currently exist for measuring glucose, but these techniques are limited by complexity of the measurement, requirement of bulky equipment, and low temporal/spatial resolution. The development of various types of biosensors (eg, electrochemical, optical sensors) for laboratory and/or clinical applications will provide new insights into the cause(s) and possible treatments of diabetes. State-of-the-art biosensors are improved by incorporating catalytic nanomaterials such as carbon nanotubes, graphene, electrospun nanofibers, and quantum dots. These nanomaterials greatly enhance biosensor performance, namely sensitivity, response time, and limit of detection. A wide range of new biosensors that incorporate nanomaterials such as lab-on-chip and nanosensor devices are currently being developed for in vivo and in vitro glucose sensing. These real-time monitoring tools represent a powerful diagnostic and monitoring tool for measuring glucose in diabetes research and point of care diagnostics. However, concerns over the possible toxicity of some nanomaterials limit the application of these devices for in vivo sensing. This review provides a general overview of the state of the art in nanomaterial-mediated biosensors for in vivo and in vitro glucose sensing, and discusses some of the challenges associated with nanomaterial toxicity. PMID:24876594

  18. Optical Fiber LSPR Biosensor Prepared by Gold Nanoparticle Assembly on Polyelectrolyte Multilayer

    PubMed Central

    Shao, Yunliang; Xu, Shuping; Zheng, Xianliang; Wang, Ye; Xu, Weiqing

    2010-01-01

    This article provides a novel method of constructing an optical fiber localized surface plasmon resonance (LSPR) biosensor. A gold nanoparticle (NP) assembled film as the sensing layer was built on the polyelectrolyte (PE) multilayer modified sidewall of an unclad optical fiber. By using a trilayer PE structure, we obtained a monodisperse gold NP assembled film. The preparation procedure for this LSPR sensor is simple and time saving. The optical fiber LSPR sensor has higher sensitivity and outstanding reproducibility. The higher anti-interference ability for response to an antibody makes it a promising method in application as a portable immuno-sensor. PMID:22319313

  19. Ultra-sensitive biosensor for K-ras gene detection using enzyme capped gold nanoparticles conjugates for signal amplification.

    PubMed

    Fang, Xian; Bai, Lijuan; Han, Xiaowei; Wang, Jiao; Shi, Anqi; Zhang, Yuzhong

    2014-09-01

    In this study, an ultra-sensitive hairpin DNA-based electrochemical DNA biosensor for K-ras gene detection is described. Gold nanoparticles (Au-NPs) and horseradish peroxidase (HRP)-streptavidin capped Au-NPs (HAS) conjugates are used for signal amplification. Initially, hairpin DNA dually labeled with thiol at its 5' end and with biotin at its 3' end is immobilized on the surface of Au-NPs modified electrode, and the hairpin DNA is in a "closed" state; hence, the HAS conjugates are shielded from being approached by the biotin due to steric hindrance. However, in the presence of target DNA, the target DNA hybridizes with the loop structure of hairpin DNA and causes conformational change, resulting in biotin forced away from the electrode surface, thereby becoming accessible for the HAS conjugates. Thus, the HAS conjugates are linked to the electrode surface via the specific interaction between biotin and streptavidin. Electrochemical detection was performed in phosphate-buffered saline (PBS) containing tetramethylbenzidine (TMB) and H2O2. Under optimal conditions, the peak current differences (ΔI) are linear with the target DNA in the range from 0.1 fM to 1 nM with a detection limit of 0.035 fM. Furthermore, this biosensor also demonstrates its excellent specificity for single-base mismatched DNA. PMID:24939462

  20. 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

  1. MRI Biosensors: A Short Primer

    PubMed Central

    Louie, Angelique

    2013-01-01

    Interest in Magnetic Resonance Imaging (MRI) contrast agents for molecular imaging of biological function experienced a surge of excitement approximately 20 years ago with the development of the first activatable contrast agents that could act as biosensors and turn “on” in response to a specific biological activity. This brief tutorial, based on a short course lecture from the 2011 ISMRM meeting, provides an overview of underlying principles governing the design of biosensing contrast agents. We describe mechanisms by which a magnetic resonance imaging (MRI) contrast agent can be made into a sensor for both T1 and T2 types contrast agents. Examples of biological activities that can interact with a contrast agent are discussed using specific examples from the recent literature to illustrate the primary mechanisms of action that have been utilized to achieve activation. MRI sensors for pH, ion binding, enzyme cleavage, and oxidation-reduction are presented. This article is not meant to be an exhaustive review, but an illustrative primer to explain how activation can be achieved for an MRI contrast agent. Chemical exchange saturation transfer (CEST) is not covered as these agents were covered in a separate lecture. PMID:23996662

  2. 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

  3. Single electrode biosensor for simultaneous determination of interferon gamma and lysozyme.

    PubMed

    Xia, Jianfei; Song, Daimin; Wang, Zonghua; Zhang, Feifei; Yang, Min; Gui, Rijun; Xia, Lin; Bi, Sai; Xia, Yanzhi; Li, Yanhui; Xia, Linhua

    2015-06-15

    Simultaneous detection of multiple biomarkers holds great promise for acute leukemia evaluation. Here, a novel biosensor is developed for simultaneous electrochemical detection of interferon gamma (IFN-γ) and lysozyme (Lys) based on aptamer recognition by coupling "signal-on" and "signal-off" modes. On one Au electrode, two kinds of signaling probes labeled by the thiolated ferrocene (Fc)- and methy blue (MB)- were designed to hybridize with IFN-γ and Lys aptamers respectively to form partial complementary DNA duplexes. In the presence of IFN-γ and Lys, the target-aptamer interaction led to the release of aptamer from duplex DNA structure. The single-stranded signaling probes thus suffered from the conformation changes, which resulted in the decreased (or increased) oxidation peak current of Fc (or MB) according to the "signal-off (or signal-on)" mode. Electrodes were characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under the optimized conditions, the signal changes were quantified using square wave voltammetry (SWV). This proposed biosensor for IFN-γ and Lys possessed linear detection range from 0.01 to 10 nM and 0.1 to 100 nM, with the detection limits of 1.14×10(-3) nM and 0.0164 nM, respectively. Moreover, this biosensor was readily regenerated and proved successful toward the practical analysis. The proposed strategy could provide more integrated and reliable information for acute leukemia evaluation. PMID:25558873

  4. Transcription factor-based biosensors enlightened by the analyte

    PubMed Central

    Fernandez-López, Raul; Ruiz, Raul; de la Cruz, Fernando; Moncalián, Gabriel

    2015-01-01

    Whole cell biosensors (WCBs) have multiple applications for environmental monitoring, detecting a wide range of pollutants. WCBs depend critically on the sensitivity and specificity of the transcription factor (TF) used to detect the analyte. We describe the mechanism of regulation and the structural and biochemical properties of TF families that are used, or could be used, for the development of environmental WCBs. Focusing on the chemical nature of the analyte, we review TFs that respond to aromatic compounds (XylS-AraC, XylR-NtrC, and LysR), metal ions (MerR, ArsR, DtxR, Fur, and NikR) or antibiotics (TetR and MarR). Analyzing the structural domains involved in DNA recognition, we highlight the similitudes in the DNA binding domains (DBDs) of these TF families. Opposite to DBDs, the wide range of analytes detected by TFs results in a diversity of structures at the effector binding domain. The modular architecture of TFs opens the possibility of engineering TFs with hybrid DNA and effector specificities. Yet, the lack of a crisp correlation between structural domains and specific functions makes this a challenging task. PMID:26191047

  5. Thin Hydrogel Films for Optical Biosensor Applications

    PubMed Central

    Mateescu, Anca; Wang, Yi; Dostalek, Jakub; Jonas, Ulrich

    2012-01-01

    Hydrogel materials consisting of water-swollen polymer networks exhibit a large number of specific properties highly attractive for a variety of optical biosensor applications. This properties profile embraces the aqueous swelling medium as the basis of biocompatibility, non-fouling behavior, and being not cell toxic, while providing high optical quality and transparency. The present review focuses on some of the most interesting aspects of surface-attached hydrogel films as active binding matrices in optical biosensors based on surface plasmon resonance and optical waveguide mode spectroscopy. In particular, the chemical nature, specific properties, and applications of such hydrogel surface architectures for highly sensitive affinity biosensors based on evanescent wave optics are discussed. The specific class of responsive hydrogel systems, which can change their physical state in response to externally applied stimuli, have found large interest as sophisticated materials that provide a complex behavior to hydrogel-based sensing devices. PMID:24957962

  6. Antibodies and antibody-derived analytical biosensors.

    PubMed

    Sharma, Shikha; Byrne, Hannah; O'Kennedy, Richard J

    2016-06-30

    The rapid diagnosis of many diseases and timely initiation of appropriate treatment are critical determinants that promote optimal clinical outcomes and general public health. Biosensors are now being applied for rapid diagnostics due to their capacity for point-of-care use with minimum need for operator input. Antibody-based biosensors or immunosensors have revolutionized diagnostics for the detection of a plethora of analytes such as disease markers, food and environmental contaminants, biological warfare agents and illicit drugs. Antibodies are ideal biorecognition elements that provide sensors with high specificity and sensitivity. This review describes monoclonal and recombinant antibodies and different immobilization approaches crucial for antibody utilization in biosensors. Examples of applications of a variety of antibody-based sensor formats are also described. PMID:27365031

  7. Hydrogen peroxide biosensor based on titanium oxide

    NASA Astrophysics Data System (ADS)

    Halim, Nur Hamidah Abdul; Heng, Lee Yook; Hashim, Uda

    2015-09-01

    In this work, a biosensor utilizing modified titania, TiO2 particles using aminopropyl-triethoxy-silane, (APTS) for developing hydrogen peroxide biosensor is presented. The surface of Ti-APTS particles is used as a support for hemoglobin immobilization via covalent bonding. The performance of the biosensor is determined by differential pulse voltammetry. The linear response was observed at the reduction current of redox mediator probe [FeCN6]3-/4- at potential between 0.22 V to 0.24 V. The preliminary result for electrochemistry study on this modified electrode is reported. The preliminary linear range is obtained from 1×10-2 M to 1×10-8 M.

  8. Antibodies and antibody-derived analytical biosensors

    PubMed Central

    Sharma, Shikha; Byrne, Hannah

    2016-01-01

    The rapid diagnosis of many diseases and timely initiation of appropriate treatment are critical determinants that promote optimal clinical outcomes and general public health. Biosensors are now being applied for rapid diagnostics due to their capacity for point-of-care use with minimum need for operator input. Antibody-based biosensors or immunosensors have revolutionized diagnostics for the detection of a plethora of analytes such as disease markers, food and environmental contaminants, biological warfare agents and illicit drugs. Antibodies are ideal biorecognition elements that provide sensors with high specificity and sensitivity. This review describes monoclonal and recombinant antibodies and different immobilization approaches crucial for antibody utilization in biosensors. Examples of applications of a variety of antibody-based sensor formats are also described. PMID:27365031

  9. Microfabricated glucose biosensor for culture well operation.

    PubMed

    Pemberton, R M; Cox, T; Tuffin, R; Sage, I; Drago, G A; Biddle, N; Griffiths, J; Pittson, R; Johnson, G; Xu, J; Jackson, S K; Kenna, G; Luxton, R; Hart, J P

    2013-04-15

    A water-based carbon screen-printing ink formulation, containing the redox mediator cobalt phthalocyanine (CoPC) and the enzyme glucose oxidase (GOx), was investigated for its suitability to fabricate glucose microbiosensors in a 96-well microplate format: (1) the biosensor ink was dip-coated onto a platinum (Pt) wire electrode, leading to satisfactory amperometric performance; (2) the ink was deposited onto the surface of a series of Pt microelectrodes (10-500 μm diameter) fabricated on a silicon substrate using MEMS (microelectromechanical systems) microfabrication techniques: capillary deposition proved to be successful; a Pt microdisc electrode of ≥100 μm was required for optimum biosensor performance; (3) MEMS processing was used to fabricate suitably sized metal (Pt) tracks and pads onto a silicon 96 well format base chip, and the glucose biosensor ink was screen-printed onto these pads to create glucose microbiosensors. When formed into microwells, using a 340 μl volume of buffer, the microbiosensors produced steady-state amperometric responses which showed linearity up to 5 mM glucose (CV=6% for n=5 biosensors). When coated, using an optimised protocol, with collagen in order to aid cell adhesion, the biosensors continued to show satisfactory performance in culture medium (linear range to 2 mM, dynamic range to 7 mM, CV=5.7% for n=4 biosensors). Finally, the operation of these collagen-coated microbiosensors, in 5-well 96-well format microwells, was tested using a 5-channel multipotentiostat. A relationship between amperometric response due to glucose, and cell number in the microwells, was observed. These results indicate that microphotolithography and screen-printing techniques can be combined successfully to produce microbiosensors capable of monitoring glucose metabolism in 96 well format cell cultures. The potential application areas for these microbiosensors are discussed. PMID:23265827

  10. Development of a biosensor for caffeine.

    PubMed

    Babu, V R Sarath; Patra, S; Karanth, N G; Kumar, M A; Thakur, M S

    2007-01-23

    We have utilized a microbe, which can degrade caffeine to develop an Amperometric biosensor for determination of caffeine in solutions. Whole cells of Pseudomonas alcaligenes MTCC 5264 having the capability to degrade caffeine were immobilized on a cellophane membrane with a molecular weight cut off (MWCO) of 3000-6000 by covalent crosslinking method using glutaraledhyde as the bifunctional crosslinking agent and gelatin as the protein based stabilizing agent (PBSA). The biosensor system was able to detect caffeine in solution over a concentration range of 0.1 to 1 mg mL(-1). With read-times as short as 3 min, this caffeine biosensor acts as a rapid analysis system for caffeine in solutions. Interestingly, successful isolation and immobilization of caffeine degrading bacteria for the analysis of caffeine described here was enabled by a novel selection strategy that incorporated isolation of caffeine degrading bacteria capable of utilizing caffeine as the sole source of carbon and nitrogen from soils and induction of caffeine degrading capacity in bacteria for the development of the biosensor. This biosensor is highly specific for caffeine and response to interfering compounds such as theophylline, theobromine, paraxanthine, other methyl xanthines and sugars was found to be negligible. Although a few biosensing methods for caffeine are reported, they have limitations in application for commercial samples. The development and application of new caffeine detection methods remains an active area of investigation, particularly in food and clinical chemistry. The optimum pH and temperature of measurement were 6.8 and 30+/-2 degrees C, respectively. Interference in analysis of caffeine due to different substrates was observed but was not considerable. Caffeine content of commercial samples of instant tea and coffee was analyzed by the biosensor and the results compared well with HPLC analysis. PMID:17386510

  11. An evaluation of hybridization kinetics in biosensors using a single-fractal analysis.

    PubMed

    Vontel, S; Ramakrishnan, A; Sadana, A

    2000-04-01

    The diffusion-limited hybridization kinetics of analyte in solution to receptor immobilized on a biosensor or immunosensor surface is analysed within a fractal framework. The data may be analysed by a single-fractal analysis. This was indicated by the regression analysis provided by Sigmaplot [Sigmaplot Users Manual (1993) Jandel Scientific, San Rafael, CA]. It is of interest to note that the binding-rate coefficient and the fractal dimension both exhibit changes in the same and in the opposite directions for the single example presented in each case. The binding-rate coefficient(s) expressions developed as a function of the analyte (DNA) concentration in solution and the fractal dimension are of particular value, since they provide a means to better control biosensor or immunosensor performance and provide physical insights into the hybridization process. PMID:10744961

  12. Quantification of the surface stress in microcantilever biosensors: revisiting Stoney's equation.

    PubMed

    Tamayo, Javier; Ruz, Jose J; Pini, Valerio; Kosaka, Priscila; Calleja, Montserrat

    2012-11-30

    Microcantilever biosensors in the static operation mode translate molecular recognition into a surface stress signal. Surface stress is derived from the nanomechanical cantilever bending by applying Stoney's equation, derived more than 100 years ago. This equation ignores the clamping effect on the cantilever deformation, which induces significant errors in the quantification of the biosensing response. This leads to discrepancies in the surface stress induced by biomolecular interactions in measurements with cantilevers with different sizes and geometries. So far, more accurate solutions have been precluded by the formidable complexity of the theoretical problem that involves solving the two-dimensional biharmonic equation. In this paper, we present an accurate and simple analytical expression to quantify the response of microcantilever biosensors. The equation exhibits an excellent agreement with finite element simulations and DNA immobilization experiments on gold-coated microcantilevers. PMID:23103805

  13. Thermoresponsive magnetic nano-biosensors for rapid measurements of inorganic arsenic and cadmium.

    PubMed

    Siddiki, Mohammad Shohel Rana; Shimoaoki, Shun; Ueda, Shunsaku; Maeda, Isamu

    2012-01-01

    Green fluorescent protein-tagged sensor proteins, ArsR-GFP and CadC-GFP, have been produced as biosensors for simple and low-cost quantification of As(III) or Cd(II). In this study, the sensor protein-promoter DNA complexes were reconstructed on the surfaces of magnetic particles of different sizes. After the surface modification all the particles could be attracted by magnets, and released different amounts of GFP-tagged protein, according to the metal concentrations within 5 min, which caused significant increases in fluorescence. A detection limit of 1 µg/L for As(III) and Cd(II) in purified water was obtained only with the nanoparticles exhibiting enough magnetization after heat treatment for 1 min. Therefore, thermoresponsive magnetic nano-biosensors offer great advantages of rapidity and sensitivity for the measurement of the toxic metals in drinking water. PMID:23202034

  14. A novel firefly luciferase biosensor enhances the detection of apoptosis induced by ESAT-6 family proteins of Mycobacterium tuberculosis

    SciTech Connect

    Shi, Junwei; Zhang, Huan; Fang, Liurong; Xi, Yongqiang; Zhou, Yanrong; Luo, Rui; Wang, Dang Xiao, Shaobo; Chen, Huanchun

    2014-10-03

    Highlights: • We developed a novel firefly luciferase based biosensor to detect apoptosis. • The novel biosensor 233-DnaE-DEVDG was reliable, sensitive and convenient. • 233-DnaE-DEVDG faithfully indicated ESAT-6 family proteins of Mycobacterium tuberculosis induced apoptosis. • EsxA, esxT and esxL in ESAT-6 family proteins induced apoptosis. • Activation of nuclear factor-κB (NF-κB) participated in esxT-induced apoptosis. - Abstract: The activation of caspase-3 is a key surrogate marker for detecting apoptosis. To quantitate caspase-3 activity, we constructed a biosensor comprising a recombinant firefly luciferase containing a caspase-3 cleavage site. When apoptosis was induced, caspase-3 cleavage of the biosensor activated firefly luciferase by a factor greater than 25. The assay conveniently detected apoptosis in real time, indicating that it will facilitate drug discovery. We screened ESAT-6 family proteins of Mycobacterium tuberculosis and found that esxA, esxT and esxL induced apoptosis. Further, activation of nuclear factor-κB (NF-κB) and the NF-κB-regulated genes encoding tumor necrosis factor-α (TNF-α) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) participated in esxT-induced apoptosis. We conclude that this assay is useful for high-throughput screening to identify and characterize proteins and drugs that regulate apoptosis.

  15. Biosensor technology for pesticides--a review.

    PubMed

    Verma, Neelam; Bhardwaj, Atul

    2015-03-01

    Pesticides, due to their lucrative outcomes, are majorly implicated in agricultural fields for crop production enhancement. Due to their pest removal properties, pesticides of various classes have been designed to persist in the environment over a longer duration after their application to achieve maximum effectiveness. Apart from their recalcitrant structure and agricultural benefits, pesticides also impose acute toxicological effects onto the other various life forms. Their accumulation in the living system may prove to be detrimental if established in higher concentrations. Thus, their prompt and accurate analysis is a crucial matter of concern. Conventional techniques like chromatographic techniques (HPLC, GC, etc.) used for pesticides detection are associated with various limitations like stumpy sensitivity and efficiency, time consumption, laboriousity, requirement of expensive equipments and highly trained technicians, and many more. So there is a need to recruit the methods which can detect these neurotoxic compounds sensitively, selectively, rapidly, and easily in the field. Present work is a brief review of the pesticide effects, their current usage scenario, permissible limits in various food stuffs and 21st century advancements of biosensor technology for pesticide detection. Due to their exceptional performance capabilities, easiness in operation and on-site working, numerous biosensors have been developed for bio-monitoring of various environmental samples for pesticide evaluation immensely throughout the globe. Till date, based on sensing element (enzyme based, antibody based, etc.) and type of detection method used (Electrochemical, optical, and piezoelectric, etc.), a number of biosensors have been developed for pesticide detection. In present communication, authors have summarized 21st century's approaches of biosensor technology for pesticide detection such as enzyme-based biosensors, immunosensors, aptamers, molecularly imprinted polymers, and

  16. Assembling Amperometric Biosensors for Clinical Diagnostics

    PubMed Central

    Belluzo, María Soledad; Ribone, María Élida; Lagier, Claudia Marina

    2008-01-01

    Clinical diagnosis and disease prevention routinely require the assessment of species determined by chemical analysis. Biosensor technology offers several benefits over conventional diagnostic analysis. They include simplicity of use, specificity for the target analyte, speed to arise to a result, capability for continuous monitoring and multiplexing, together with the potentiality of coupling to low-cost, portable instrumentation. This work focuses on the basic lines of decisions when designing electron-transfer-based biosensors for clinical analysis, with emphasis on the strategies currently used to improve the device performance, the present status of amperometric electrodes for biomedicine, and the trends and challenges envisaged for the near future.

  17. Fluorescent labels in biosensors for pathogen detection.

    PubMed

    Li, Bianmiao; Yu, Qiaoling; Duan, Yixiang

    2015-03-01

    Infectious diseases caused by pathogens have become a life-threatening problem for millions of people around the world in recent years. Therefore, the need of efficient, fast, low-cost and user-friendly biosensing systems to monitor pathogen has increased enormously in the last few years. This paper presents an overview of different fluorescent labels and the utilization of fluorescence-based biosensor techniques for rapid, direct, sensitive and real-time identification of bacteria. In these biosensors, organic dyes, nanomaterials and rare-earth elements are playing an increasing role in the design of biosensing systems with an interest for applications in bacterial analysis. PMID:23886349

  18. Mathematical Model of the Biosensors Acting in a Trigger Mode

    PubMed Central

    Baronas, Romas; Kulys, Juozas; Ivanauskas, Feliksas

    2004-01-01

    A mathematical model of biosensors acting in a trigger mode has been developed. One type of the biosensors utilized a trigger enzymatic reaction followed by the cyclic enzymatic and electrochemical conversion of the product (CCE scheme). Other biosensors used the enzymatic trigger reaction followed by the electrochemical and enzymatic product cyclic conversion (CEC scheme). The models were based on diffusion equations containing a non-linear term related to Michaelis-Menten kinetics of the enzymatic reactions. The digital simulation was carried out using the finite difference technique. The influence of the substrate concentration, the maximal enzymatic rate as well as the membrane thickness on the biosensor response was investigated. The numerical experiments demonstrated a significant gain (up to dozens of times) in biosensor sensitivity when the biosensor response was under diffusion control. In the case of significant signal amplification, the response time with triggering was up to several times longer than that of the biosensor without triggering.

  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. Cell buffer with built-in test

    NASA Technical Reports Server (NTRS)

    Ott, William E. (Inventor)

    2004-01-01

    A cell buffer with built-in testing mechanism is provided. The cell buffer provides the ability to measure voltage provided by a power cell. The testing mechanism provides the ability to test whether the cell buffer is functioning properly and thus providing an accurate voltage measurement. The testing mechanism includes a test signal-provider to provide a test signal to the cell buffer. During normal operation, the test signal is disabled and the cell buffer operates normally. During testing, the test signal is enabled and changes the output of the cell buffer in a defined way. The change in the cell buffer output can then be monitored to determine if the cell buffer is functioning correctly. Specifically, if the voltage output of the cell buffer changes in a way that corresponds to the provided test signal, then the functioning of the cell buffer is confirmed. If the voltage output of the cell buffer does not change correctly, then the cell buffer is known not to be operating correctly. Thus, the built in testing mechanism provides the ability to quickly and accurately determine if the cell buffer is operating correctly. Furthermore, the testing mechanism provides this functionality without requiring excessive device size and complexity.

  1. DNA attachment to nanocrystalline diamond films

    NASA Astrophysics Data System (ADS)

    Wenmackers, S.; Christiaens, P.; Daenen, M.; Haenen, K.; Nesládek, M.; van Deven, M.; Vermeeren, V.; Michiels, L.; Ameloot, M.; Wagner, P.

    2005-09-01

    A biochemical method to immobilize DNA on synthetic diamond for biosensor applications is developed. Nanocrystalline diamond is grown using microwave plasma-enhanced chemical vapour deposition. On the hydrogen-terminated surface 10-undecenoic acid is tethered photochemically under 254 nm illumination, followed by 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide crosslinker-mediated attachment of amino modified DNA. The attachment is functionally confirmed by comparison of supernatant fluorescence and gel electrophoresis. The linking procedure allowed for 35 denaturation and rehybridisation steps.

  2. Biosensor Systems for Homeland Security

    SciTech Connect

    Bruckner-Lea, Cindy J.

    2004-05-30

    The detection of biological agents is important to minimize the effects of pathogens that can harm people, livestock, or plants. In addition to pathogens distributed by man, there is a need to detect natural outbreaks. Recent outbreaks of SARS, mad cow disease, pathogenic E. coli and Salmonella, as well as the discovery of letters filled with anthrax spores have highlighted the need for biosensor systems to aid in prevention, early warning, response, and recovery. Rapid detection can be used to prevent exposure; and detection on a longer timescale can be used to minimize exposure, define treatment, and determine whether contaminated areas are clean enough for reuse. The common types of biological agents of concern include bacteria, spores, and viruses (Figure 1). From a chemist’s point of view, pathogens are essentially complex packages of chemicals that are assembled into organized packages with somewhat predictable physical characteristics such as size and shape. Pathogen detection methods can be divided into three general approaches: selective detection methods for specific identification such as nucleic acid analysis and structural recognition, semi-selective methods for broad-spectrum detection (e.g. physical properties, metabolites, lipids), and function-based methods (e.g. effect of the pathogen on organisms, tissues, or cells). The requirements for biodetection systems depend upon the application. While detect to warn sensors may require rapid detection on the order one minute, detection times of many minutes or hours may be suitable for determining appropriate treatments or for forensic analysis. Of course ideal sensor systems will meet the needs of many applications, and will be sensitive, selective, rapid, and simultaneously detect all agents of concern. They will also be reliable with essentially no false negatives or false positives, small, easy to use, and low cost with minimal consumables.

  3. Synthesis and biosensor application of Ag@Au bimetallic nanoparticles based on localized surface plasmon resonance

    NASA Astrophysics Data System (ADS)

    Ghodselahi, T.; Arsalani, S.; Neishaboorynejad, T.

    2014-05-01

    This work demonstrates a simple method for synthesizing gold-silver bimetallic nanoparticles (Ag@Au BNPs). Ag@Au BNPs on the carbon thin film are prepared by co-deposition of RF-sputtering and RF-PECVD using acetylene gas and gold-silver target. X-ray diffraction analysis indicates that Au and Ag NPs with FCC crystal structure are formed in our samples. From AFM image and data, average particles size of gold and silver are estimated to be about 5 and 8 nm, respectively. XRD profile and localized surface plasmon resonance (LSPR) spectroscopy indicate that Ag NPs in Ag@Au BNPs composite have a more chemical activity with respect to bare Ag NPs. Biosensor application of Ag@Au BNPs without probe immobilization is introduced too. The change in LSPR absorption peak of Ag@Au BNPs in presence of DNA primer decamer (ten-deoxycytosine) at fM concentrations is investigated. The LSPR absorption peak of Au NPs has a blue shift and the LSPR absorption peak of Ag NPs has a red shift by addition of DNA primer and under DNA exposure up to 1 h. Our sample shows a good response to low concentration of DNA and has a short response time. Both of these are prerequisite for applying this sample as LSPR biosensor chip.

  4. Doublet III beamline: as-built

    SciTech Connect

    Harder, C.R.; Holland, M.M.; Parker, J.W.; Gunn, J.; Resnick, L.

    1980-03-01

    In order to fully exploit Doublet III capabilities and to study new plasma physics regimes, a Neutral Beam Injector System has been constructed. Initially, a two beamline system will supply 7 MW of heat to the plasma. The system is currently being expanded to inject approx. 20 MW of power (6 beamlines). Each beamline is equipped with two Lawrence Berkeley Laboratory type rectangular ion sources with 10 cm x 40 cm extraction grids. These sources will accelerate hydrogen ions to 80 keV, with extracted beam currents in excess of 80 A per source expected. The first completed source is currently being tested and conditioned on the High Voltage Test Stand at Lawrence Livermore Laboratory. This paper pictorially reviews the as-built Doublet III neutral beamline with emphasis on component relation and configuration relative to spatial and source imposed design constraints.

  5. Tester-assisted built in test

    NASA Astrophysics Data System (ADS)

    Guntheroth, Kurt

    It is noted that board makers invest considerable time and money writing extensive self-tests and that this investment can be multiplied by selecting ATE (automatic test equipment) that complements and extends the power of the self-test. The tester can diagnose boards in situations where a fault prevents the self-test from running. If the tester monitors such resources as processor, memory, and I/O, confidence in test results is improved. The tester can be used during development of the self-test and to turn on prototypes before the self-test is complete. The author argues that emulative functional testers outperform other types of ATE on boards with BIST (built-in self-test) and lists features of emulative functional testers that are most important to users of BIST.

  6. The Large Built Water Clock Of Amphiaraeion.

    NASA Astrophysics Data System (ADS)

    Theodossiou, E.; Katsiotis, M.; Manimanis, V. N.; Mantarakis, P.

    A very well preserved ancient water clock was discovered during excavations at the Amphiaraeion, in Oropos, Greece. The Amphiaraeion, a famous religious and oracle center of the deified healer Amphiaraus, was active from the pre-classic period until the replacement of the ancient religion by Christianity in the 5th Century A.D.. The foretelling was supposedly done through dreams sent by the god to the believers sleeping in a special gallery. In these dreams the god suggesting to them the therapy for their illness or the solution to their problems. The patients, then threw coins into a spring of the sanctuary. In such a place, the measurement of time was a necessity. Therefore, time was kept with both a conical sundial and a water clock in the form of a fountain. According to archeologists, the large built structure that measured the time for the sanctuary dates from the 4th Century B.C.

  7. Microbiosensors based on DNA modified single-walled carbon nanotube and Pt black nanocomposites.

    PubMed

    Shi, Jin; Cha, Tae-Gon; Claussen, Jonathan C; Diggs, Alfred R; Choi, Jong Hyun; Porterfield, D Marshall

    2011-12-01

    Glucose and ATP biosensors have important applications in diagnostics and research. Biosensors based on conventional materials suffer from low sensitivity and low spatial resolution. Our previous work has shown that combining single-walled carbon nanotubes (SWCNTs) with Pt nanoparticles can significantly enhance the performance of electrochemical biosensors. The immobilization of SWCNTs on biosensors remains challenging due to the aqueous insolubility originating from van der Waals forces. In this study, we used single-stranded DNA (ssDNA) to modify SWCNTs to increase solubility in water. This allowed us to explore new schemes of combining ssDNA-SWCNT and Pt black in aqueous media systems. The result is a nanocomposite with enhanced biosensor performance. The surface morphology, electroactive surface area, and electrocatalytic performance of different fabrication protocols were studied and compared. The ssDNA-SWCNT/Pt black nanocomposite constructed by a layered scheme proved most effective in terms of biosensor activity. The key feature of this protocol is the exploitation of ssDNA-SWCNTs as molecular templates for Pt black electrodeposition. The glucose and ATP microbiosensors fabricated on this platform exhibited high sensitivity (817.3 nA/mM and 45.6 nA/mM, respectively), wide linear range (up to 7 mM and 510 μM), low limit of detection (1 μM and 2 μM) and desirable selectivity. This work is significant to biosensor development because this is the first demonstration of ssDNA-SWCNT/Pt black nanocomposite as a platform for constructing both single-enzyme and multi-enzyme biosensors for physiological applications. PMID:21858297

  8. Application of the magnetoimpedance to biosensors

    NASA Astrophysics Data System (ADS)

    Song, Hoon; Park, Duck-gun

    2012-11-01

    A prototype of a giant magnetoimpedance (GMI) biosensor was designed and tested using an amorphous ribbon. The GMI response was measured with a suspension containing a CM01N/9077 bead from Bangs Laboratories. This bead was coated with streptavidin so that it could be used as biomolecular label with a specific antibody. The GMI change caused by the presence of the specific antibody was measured with a prototype biosensor designed with a measuring cell containing an amorphous CoFeBSi ribbon. The GMI was measured in a range of current frequencies from 0.1 to 10 MHz and at intensities of Irms = 5 mA. Commercial Bangs Laboratories beads were supplied as a suspension in phosphate-buffered saline. A maximum difference of 5% in the GMI ratio measured with and without magnetic beads was obtained at a frequency of 1 MHz and Irms = 5 mA. As some potential applications for the GMI biosensor and further directions, a magnetoimpedance biosensor for detecting the presence of magnetic nanoparticles after introducing a protein on the surface of the sensor has been developed and tested for a model system by using human P21 protein (antibody and antigen) and nonspecific magnetic nanoparticles.

  9. Methods for using redox liposome biosensors

    DOEpatents

    Cheng, Quan; Stevens, Raymond C.

    2002-01-01

    The present invention provides methods and compositions for detecting the presence of biologically-important analytes by using redox liposome biosensors. In particular, the present invention provides liposome/sol-gel electrodes suitable for the detection of a wide variety of organic molecules, including but not limited to bacterial toxins.

  10. Boar taint detection using parasitoid biosensors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To evaluate the potential for a non-stinging wasp to be used as a biosensor in the pig industry, we trained wasps to 3 individual chemicals associated with boar taint. Training consisted of presenting the odors to hungry wasps while they were feeding on sugar. This associates the chemical with a fo...

  11. Clinical Assessment Applications of Ambulatory Biosensors

    ERIC Educational Resources Information Center

    Haynes, Stephen N.; Yoshioka, Dawn T.

    2007-01-01

    Ambulatory biosensor assessment includes a diverse set of rapidly developing and increasingly technologically sophisticated strategies to acquire minimally disruptive measures of physiological and motor variables of persons in their natural environments. Numerous studies have measured cardiovascular variables, physical activity, and biochemicals…

  12. Microbial Biosensors for Selective Detection of Disaccharides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seven microbial strains were screened for their ability to detect disaccharides as components of Clark-type oxygen biosensors. Sensors responded to varying degrees to maltose, cellobiose, sucrose, and melibiose, but none responded strongly to lactose. Although microbial sensors are relatively nons...

  13. [Quantitative analysis of calibration dependence of biosensors].

    PubMed

    Borisov, I A; Lobanov, A V; Reshetilov, A N; Kurganov, B I

    2000-01-01

    Three-parameter Hill's equation, which is used in enzyme kinetics, was shown to applicable to calibration curves of both potentiometric (glucose, pesticides, urea, etc.) and amperometric (surfactants, biphenyl, etc.) biosensors. Possible causes of errors of analyte concentration measurements are discussed. PMID:10867941

  14. Cell-based biosensors in clinical chemistry.

    PubMed

    Kintzios, Spiridon E

    2007-10-01

    Cell-based biosensors represent the next revolution in medical diagnostics, offering a number of significant advantages, such as high speed, portability and low cost. The present review focuses on the most successful technologies used for the detection of ultra-low concentrations of bioactive analytes (such as metabolic markers and pathogens) in clinical samples. PMID:17979804

  15. Development and Applications of Portable Biosensors.

    PubMed

    Srinivasan, Balaji; Tung, Steve

    2015-08-01

    The significance of microfluidics-based and microelectromechanical systems-based biosensors has been widely acknowledged, and many reviews have explored their potential applications in clinical diagnostics, personalized medicine, global health, drug discovery, food safety, and forensics. Because health care costs are increasing, there is an increasing need to remotely monitor the health condition of patients by point-of-care-testing. The demand for biosensors for detection of biological warfare agents has increased, and research is focused on ways of producing small portable devices that would allow fast, accurate, and on-site detection. In the past decade, the demand for rapid and accurate on-site detection of plant disease diagnosis has increased due to emerging pathogens with resistance to pesticides, increased human mobility, and regulations limiting the application of toxic chemicals to prevent spread of diseases. The portability of biosensors for on-site diagnosis is limited due to various issues, including sample preparation techniques, fluid-handling techniques, the limited lifetime of biological reagents, device packaging, integrating electronics for data collection/analysis, and the requirement of external accessories and power. Many microfluidic, electronic, and biological design strategies, such as handling liquids in biosensors without pumps/valves, the application of droplet-based microfluidics, paper-based microfluidic devices, and wireless networking capabilities for data transmission, are being explored. PMID:25878051

  16. Biosensors for Whole-Cell Bacterial Detection

    PubMed Central

    Rushworth, Jo V.; Hirst, Natalie A.; Millner, Paul A.

    2014-01-01

    SUMMARY Bacterial pathogens are important targets for detection and identification in medicine, food safety, public health, and security. Bacterial infection is a common cause of morbidity and mortality worldwide. In spite of the availability of antibiotics, these infections are often misdiagnosed or there is an unacceptable delay in diagnosis. Current methods of bacterial detection rely upon laboratory-based techniques such as cell culture, microscopic analysis, and biochemical assays. These procedures are time-consuming and costly and require specialist equipment and trained users. Portable stand-alone biosensors can facilitate rapid detection and diagnosis at the point of care. Biosensors will be particularly useful where a clear diagnosis informs treatment, in critical illness (e.g., meningitis) or to prevent further disease spread (e.g., in case of food-borne pathogens or sexually transmitted diseases). Detection of bacteria is also becoming increasingly important in antibioterrorism measures (e.g., anthrax detection). In this review, we discuss recent progress in the use of biosensors for the detection of whole bacterial cells for sensitive and earlier identification of bacteria without the need for sample processing. There is a particular focus on electrochemical biosensors, especially impedance-based systems, as these present key advantages in terms of ease of miniaturization, lack of reagents, sensitivity, and low cost. PMID:24982325

  17. Fiber optic biosensor using aptamer as receptors

    NASA Astrophysics Data System (ADS)

    Yu, Shuqin; Cai, Xiaokun; Tan, Xianglin; Zhu, Yexiang; Lu, Bin

    2001-09-01

    Reagentless biosensor that can directly transducer molecular recognition to optical signal should potentiate the development of sensor array fora wide variety of analytes. Nucleic acid aptamer can bind ligand tightly and specifically with conformational change of aptamer, and can be used as a receptor in biosensor. We have therefore developed a fiber-optic biosensor by aptamer connected with molecular beacon. Molecular beacons consist of an oligonucleotide sequence containing complementary sequence sections at either end. These two sequence containing segments base pair with each other to form a hairpin shaped loop structure, the fluorophore and quencher were attached at 5 foot- and 3 foot-end of molecular beacon respectively. When thrombin binding to the stem-loop of molecular beacon aptamer, the pseudoknot structure was interrupted, resulting a release of fluorescence from quenching and a increase in fluorescence emission. This novel biosensor system in this project has a large potential and is specific and sensitivity. A similar strategy could be used to study other analytes such as protein and small molecules.

  18. Fiber optic-based regenerable biosensor

    DOEpatents

    Sepaniak, Michael J.; Vo-Dinh, Tuan

    1993-01-01

    A fiber optic-based regenerable biosensor. The biosensor is particularly suitable for use in microscale work in situ. In one embodiment, the biosensor comprises a reaction chamber disposed adjacent the distal end of a waveguide and adapted to receive therein a quantity of a sample containing an analyte. Leading into the chamber is a plurality of capillary conduits suitable for introducing into the chamber antibodies or other reagents suitable for selective interaction with a predetermined analyte. Following such interaction, the contents of the chamber may be subjected to an incident energy signal for developing fluorescence within the chamber that is detectable via the optical fiber and which is representative of the presence, i.e. concentration, of the selected analyte. Regeneration of the biosensor is accomplished by replacement of the reagents and/or the analyte, or a combination of these, at least in part via one or more of the capillary conduits. The capillary conduits extend from their respective terminal ends that are in fluid communication with the chamber, away from the chamber to respective location(s) remote from the chamber thereby permitting in situ location of the chamber and remote manipulation and/or analysis of the activity with the chamber.

  19. Photocatalytic oxide films in the built environment

    NASA Astrophysics Data System (ADS)

    Österlund, Lars; Topalian, Zareh

    2014-11-01

    The possibility to increase human comfort in buildings is a powerful driving force for the introduction of new technology. Among other things our sense of comfort depends on air quality, temperature, lighting level, and the possibility of having visual contact between indoors and outdoors. Indeed there is an intimate connection between energy, comfort, and health issues in the built environment, leading to a need for intelligent building materials and green architecture. Photocatalytic materials can be applied as coatings, filters, and be embedded in building materials to provide self-cleaning, antibacterial, air cleaning, deodorizing, and water cleaning functions utilizing either solar light or artificial illumination sources - either already present in buildings, or by purposefully designed luminaries. Huge improvements in indoor comfort can thus be made, and also alleviate negative health effects associated with buildings, such as the sick-house syndrome. At the same time huge cost savings can be made by reducing maintenance costs. Photocatalytic oxides can be chemically modified by changing their acid-base surface properties, which can be used to overcome deactivation problems commonly encountered for TiO2 in air cleaning applications. In addition, the wetting properties of oxides can be tailored by surface chemical modifications and thus be made e.g. oleophobic and water repellent. Here we show results of surface acid modified TiO2 coatings on various substrates by means of photo-fixation of surface sulfate species by a method invented in our group. In particular, we show that such surface treatments of photocatalytic concrete made by mixing TiO2 nanoparticles in reactive concrete powders result in concrete surfaces with beneficial self-cleaning properties. We propose that such approaches are feasible for a number of applications in the built environment, including glass, tiles, sheet metals, plastics, etc.

  20. 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

  1. Recent Advances in Biosensor Technology for Potential Applications - An Overview.

    PubMed

    Vigneshvar, S; Sudhakumari, C C; Senthilkumaran, Balasubramanian; Prakash, Hridayesh

    2016-01-01

    Imperative utilization of biosensors has acquired paramount importance in the field of drug discovery, biomedicine, food safety standards, defense, security, and environmental monitoring. This has led to the invention of precise and powerful analytical tools using biological sensing element as biosensor. Glucometers utilizing the strategy of electrochemical detection of oxygen or hydrogen peroxide using immobilized glucose oxidase electrode seeded the discovery of biosensors. Recent advances in biological techniques and instrumentation involving fluorescence tag to nanomaterials have increased the sensitive limit of biosensors. Use of aptamers or nucleotides, affibodies, peptide arrays, and molecule imprinted polymers provide tools to develop innovative biosensors over classical methods. Integrated approaches provided a better perspective for developing specific and sensitive biosensors with high regenerative potentials. Various biosensors ranging from nanomaterials, polymers to microbes have wider potential applications. It is quite important to integrate multifaceted approaches to design biosensors that have the potential for diverse usage. In light of this, this review provides an overview of different types of biosensors being used ranging from electrochemical, fluorescence tagged, nanomaterials, silica or quartz, and microbes for various biomedical and environmental applications with future outlook of biosensor technology. PMID:26909346

  2. 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.

  3. A scalable neuristor built with Mott memristors

    NASA Astrophysics Data System (ADS)

    Pickett, Matthew D.; Medeiros-Ribeiro, Gilberto; Williams, R. Stanley

    2013-02-01

    The Hodgkin-Huxley model for action potential generation in biological axons is central for understanding the computational capability of the nervous system and emulating its functionality. Owing to the historical success of silicon complementary metal-oxide-semiconductors, spike-based computing is primarily confined to software simulations and specialized analogue metal-oxide-semiconductor field-effect transistor circuits. However, there is interest in constructing physical systems that emulate biological functionality more directly, with the goal of improving efficiency and scale. The neuristor was proposed as an electronic device with properties similar to the Hodgkin-Huxley axon, but previous implementations were not scalable. Here we demonstrate a neuristor built using two nanoscale Mott memristors, dynamical devices that exhibit transient memory and negative differential resistance arising from an insulating-to-conducting phase transition driven by Joule heating. This neuristor exhibits the important neural functions of all-or-nothing spiking with signal gain and diverse periodic spiking, using materials and structures that are amenable to extremely high-density integration with or without silicon transistors.

  4. A scalable neuristor built with Mott memristors.

    PubMed

    Pickett, Matthew D; Medeiros-Ribeiro, Gilberto; Williams, R Stanley

    2013-02-01

    The Hodgkin-Huxley model for action potential generation in biological axons is central for understanding the computational capability of the nervous system and emulating its functionality. Owing to the historical success of silicon complementary metal-oxide-semiconductors, spike-based computing is primarily confined to software simulations and specialized analogue metal-oxide-semiconductor field-effect transistor circuits. However, there is interest in constructing physical systems that emulate biological functionality more directly, with the goal of improving efficiency and scale. The neuristor was proposed as an electronic device with properties similar to the Hodgkin-Huxley axon, but previous implementations were not scalable. Here we demonstrate a neuristor built using two nanoscale Mott memristors, dynamical devices that exhibit transient memory and negative differential resistance arising from an insulating-to-conducting phase transition driven by Joule heating. This neuristor exhibits the important neural functions of all-or-nothing spiking with signal gain and diverse periodic spiking, using materials and structures that are amenable to extremely high-density integration with or without silicon transistors. PMID:23241533

  5. Modelling Amperometric Biosensors Based on Chemically Modified Electrodes

    PubMed Central

    Baronas, Romas; Kulys, Juozas

    2008-01-01

    The response of an amperometric biosensor based on a chemically modified electrode was modelled numerically. A mathematical model of the biosensor is based on a system of non-linear reaction-diffusion equations. The modelling biosensor comprises two compartments: an enzyme layer and an outer diffusion layer. In order to define the main governing parameters the corresponding dimensionless mathematical model was derived. The digital simulation was carried out using the finite difference technique. The adequacy of the model was evaluated using analytical solutions known for very specific cases of the model parameters. By changing model parameters the output results were numerically analyzed at transition and steady state conditions. The influence of the substrate and mediator concentrations as well as of the thicknesses of the enzyme and diffusion layers on the biosensor response was investigated. Calculations showed complex kinetics of the biosensor response, especially when the biosensor acts under a mixed limitation of the diffusion and the enzyme interaction with the substrate.

  6. A general strategy to construct small molecule biosensors in eukaryotes

    DOE PAGESBeta

    Feng, Justin; Jester, Benjamin W.; Tinberg, Christine E.; Mandell, Daniel J.; Antunes, Mauricio S.; Chari, Raj; Morey, Kevin J.; Rios, Xavier; Medford, June I.; Church, George M.; et al

    2015-12-29

    Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Here, we produce biosensors based on a ligand-binding domain (LBD) by using a method that, in principle, can be applied to any target molecule. The LBD is fused to either a fluorescent protein or a transcriptional activator and is destabilized by mutation such that the fusion accumulates only in cells containing the target ligand. We illustrate the power of this method by developing biosensors for digoxin and progesterone. Addition of ligand to yeast, mammalian, or plant cells expressing a biosensor activatesmore » transcription with a dynamic range of up to ~100-fold. We use the biosensors to improve the biotransformation of pregnenolone to progesterone in yeast and to regulate CRISPR activity in mammalian cells. This work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes.« less

  7. Experimental and theoretical investigation effect of flavonols antioxidants on DNA damage.

    PubMed

    Ensafi, Ali A; Heydari-Soureshjani, E; Jafari-Asl, M; Rezaei, B; Ghasemi, Jahan B; Aghaee, Elham

    2015-08-01

    A new electrochemical biosensor was developed to demonstrate the effect of Acridine Orange (AO) on DNA damage. Then, the biosensor was used to check the inhibitors effect of three flavonols antioxidants (myricetin, fisetin and kaempferol) on DNA damage. Acridine Orange (AO) was used as a damaging agent because it shows a high affinity to nucleic acid and stretch of the double helical structure of DNA. Decreasing on the oxidation signals of adenine and guanine (in the DNA) in the presence of AO were used as probes to study the antioxidants power, using DNA-modified screen printed graphene electrode (DNA/SPGE). The results of our study showed that the DNA-biosensor could be suitable biosensor to investigate the inhibitors ability of the flavonols antioxidants on the DNA damage. The linear dependency was detected in the two regions in the ranges of 1.0-15.0 and 15.0-500.0 pmol L(-1). The detection limit was found 0.5 pmol L(-1) and 0.6 pmol L(-1) for guanine and adenine, respectively. To confirm the electrochemical results, Uv-Vis and fluorescence spectroscopic methods were used too. Finally molecular dynamic (MD) simulation was performed on the structure of DNA in a water box to study any interaction between the antioxidant, AO and DNA. PMID:26320789

  8. Whole-cell detection of live lactobacillus acidophilus on aptamer-decorated porous silicon biosensors.

    PubMed

    Urmann, K; Arshavsky-Graham, S; Walter, J G; Scheper, T; Segal, E

    2016-09-21

    This work describes the design of optical aptamer-based porous silicon (PSi) biosensors for the direct capture of Lactobacillus acidophilus. Aptamers are oligonucleotides (single-stranded DNA or RNA) that can bind their targets with high affinity and specificity, making them excellent recognition elements for biosensing applications. Herein, aptamer Hemag1P, which specifically targets the important probiotic L. acidophilus, was utilized for direct bacteria capture onto oxidized PSi Fabry-Pérot thin films. Monitoring changes in the reflectivity spectrum (using reflective interferometric Fourier transform spectroscopy) allows for bacteria detection in a label-free, simple and rapid manner. The performance of the biosensor was optimized by tuning the PSi nanostructure, its optical properties, as well as the immobilization density of the aptamer. We demonstrate the high selectivity and specificity of this simple "direct-capture" biosensing scheme and show its ability to distinguish between live and dead bacteria. The resulting biosensor presents a robust and rapid method for the specific detection of live L. acidophilus at concentrations relevant for probiotic products and as low as 10(6) cells per mL. Rapid monitoring of probiotic bacteria is crucial for quality, purity and safety control as the use of probiotics in functional foods and pharmaceuticals is becoming increasingly popular. PMID:27381045

  9. Biosensor-controlled gene therapy/drug delivery with nanoparticles for nanomedicine

    NASA Astrophysics Data System (ADS)

    Prow, Tarl W.; Rose, William A.; Wang, Nan; Reece, Lisa M.; Lvov, Yuri; Leary, James F.

    2005-04-01

    Nanomedicine involves cell-by-cell regenerative medicine, either repairing cells one at a time or triggering apoptotic pathways in cells that are not repairable. Multilayered nanoparticle systems are being constructed for the targeted delivery of gene therapy to single cells. Cleavable shells containing targeting, biosensing, and gene therapeutic molecules are being constructed to direct nanoparticles to desired intracellular targets. Therapeutic gene sequences are controlled by biosensor-activated control switches to provide the proper amount of gene therapy on a single cell basis. The central idea is to set up gene therapy "nanofactories" inside single living cells. Molecular biosensors linked to these genes control their expression. Gene delivery is started in response to a biosensor detected problem; gene delivery is halted when the cell response indicates that more gene therapy is not needed. Cell targeting of nanoparticles, both nanocrystals and nanocapsules, has been tested by a combination of fluorescent tracking dyes, fluorescence microscopy and flow cytometry. Intracellular targeting has been tested by confocal microscopy. Successful gene delivery has been visualized by use of GFP reporter sequences. DNA tethering techniques were used to increase the level of expression of these genes. Integrated nanomedical systems are being designed, constructed, and tested in-vitro, ex-vivo, and in small animals. While still in its infancy, nanomedicine represents a paradigm shift in thinking-from destruction of injured cells by surgery, radiation, chemotherapy to cell-by-cell repair within an organ and destruction of non-repairable cells by natural apoptosis.

  10. 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

  11. Quenching action of monofunctional sulfur mustard on chlorophyll fluorescence: towards an ultrasensitive biosensor.

    PubMed

    Kaur, Simerjit; Singh, Minni; Flora, Swaran Jeet Singh

    2013-11-01

    An ultrasensitive fluorimetric biosensor for the detection of chemical warfare agent sulfur mustard (SM) was developed using its monofunctional analogue. SM is a vesicant and a potent chemical threat owing to its direct toxic effects on eyes, lungs, skin and DNA. This work investigates the quenching action of the analyte on chlorophyll fluorescence as elucidated by nuclear magnetic resonance, Fourier transform infrared spectroscopy and mass spectrometry studies suggesting the electrophilic attack of carbonium ion on nitrogens of the porphyrin moiety of chlorophyll. The properties of immobilisation matrix were optimised and scanning electron microscope observations confirmed improvement in pore size of sol-gels by addition of 32 % (v/v) glycerol, a feature enabling enhanced sensitivity towards the analyte. Chlorophyll embedded sol-gel was treated with increasing concentrations of monofunctional SM and the corresponding drop in maximum fluorescence intensity as measured by emission at 673 nm was observed, which varied linearly and had a detection limit of 7.68 × 10(-16) M. The biosensor was found to be 6 orders of magnitude more sensitive than the glass microfibre-based disc biosensor previously reported by us. PMID:23955347

  12. Development of an RNA-based theophylline-specific microarray biosensor

    NASA Astrophysics Data System (ADS)

    Jordan, Katherine M.

    We are developing an extremely sensitive and compact biosensor that is adaptable to a variety of target analytes. Hammerhead ribozymes have been engineered such that they rearrange from a catalytically inactive to an active conformation upon binding to a target molecule. A donor-acceptor fluorophore pair is coupled to the substrate RNA of such an aptamer, to form a complex referred to as an aptazyme, to monitor real-time cleavage activity in a fluid environment. The fluorophores interact by fluorescence resonance energy transfer (FRET) until binding of the target molecule, when the FRET signal breaks down as the substrate is cleaved and the products dissociate. FRET assays with immobilized aptazymes and using total internal reflection fluorescence (TIRF) microscopy on the single-molecule scale are presented showing an enhancement of substrate cleavage in the presence of theophylline over background. The aptazyme is hybridized onto a DNA microarray and incorporated into a chip specifically designed to allow for measurement in a controlled fluid environment. The use of these microarrays allows for either one spot, or a series of spots, to be addressed independently within the biosensor. This allows for multiple analytes to be tested simultaneously. An enhancement in the substrate cleavage is again observed in the presence of theophylline. Results are presented toward the characterization of a theophylline-specific aptamer-based biosensor using this RNA microarray platform and analogous measurement techniques.

  13. New approaches for carbon nanotubes-based biosensors and their application to cell culture monitoring.

    PubMed

    Boero, Cristina; Olivo, Jacopo; De Micheli, Giovanni; Carrara, Sandro

    2012-10-01

    Amperometric biosensors are complex systems and they require a combination of technologies for their development. The aim of the present work is to propose a new approach in order to develop nanostructured biosensors for the real-time detection of multiple metabolites in cell culture flasks. The fabrication of five Au working electrodes onto silicon substrate is achieved with CMOS compatible microtechnology. Each working electrode presents an area of 0.25 mm², so structuration with carbon nanotubes and specific functionalization are carried out by using spotting technology, originally developed for microarrays and DNA printing. The electrodes are characterized by cyclic voltammetry and compared with commercially available screen-printed electrodes. Measurements are carried out under flow conditions, so a simple fluidic system is developed to guarantee a continuous flow next to the electrodes. The working electrodes are functionalized with different enzymes and calibrated for the real-time detection of glucose, lactate, and glutamate. Finally, some tests are performed on surnatant conditioned medium sampled from neuroblastoma cells (NG-108 cell line) to detect glucose and lactate concentration after 72 hours of cultivation. The developed biosensor for real-time and online detection of multiple metabolites shows very promising results towards circuits and systems for cell culture monitoring. PMID:23853234

  14. Microfluidic-integrated DNA nanobiosensors.

    PubMed

    Ansari, M I Haque; Hassan, Shabir; Qurashi, Ahsanulhaq; Khanday, Firdous Ahmad

    2016-11-15

    Over the last few decades, an increased demand has emerged for integrating biosensors with microfluidic- and nanofluidic-based lab-on-chip (LOC) devices for point-of-care (POC) diagnostics, in the medical industry and environmental monitoring of pathogenic threat agents. Such a merger of microfluidics with biosensing technologies allows for the precise control of volumes, as low as one nanolitre and the integration of various types of bioassays on a single miniaturized platform. This integration offers several favorable advantages, such as low reagent consumption, automation of sample preparation, reduction in processing time, low cost analysis, minimal handling of hazardous materials, high detection accuracy, portability and disposability. This review provides a synopsis of the most recent developments in the microfluidic-integrated biosensing field by delineating the fundamental theory of microfluidics, fabrication techniques and a detailed account of the various transduction methods that are employed. Lastly, the review discusses state-of-the-art DNA biosensors with a focus on optical DNA biosensors. PMID:27179566

  15. Life Redefined: Microbes Built with Arsenic

    SciTech Connect

    Webb, Sam

    2011-03-22

    Life can survive in many harsh environments, from extreme heat to the presence of deadly chemicals. However, life as we know it has always been based on the same six elements -- carbon, oxygen, nitrogen, hydrogen, sulfur and phosphorus. Now it appears that even this rule has an exception. In the saline and poisonous environment of Mono Lake, researchers have found a bacterium that can grow by incorporating arsenic into its structure in place of phosphorus. X-ray images taken at SLAC's synchrotron light source reveal that this microbe may even use arsenic as a building block for DNA. Please join us as we describe this discovery, which rewrites the textbook description of how living cells work.

  16. DETAIL OF TYPICAL GIRDER SPAN, PIER NUMBER 5 (BUILT IN ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    DETAIL OF TYPICAL GIRDER SPAN, PIER NUMBER 5 (BUILT IN 1922) AND PIER NUMBER 6 (BUILT IN 1929). VIEW TO NORTH. - Hassayampa Bridge, Spanning Hassayampa River at old U.S. Highway 80, Arlington, Maricopa County, AZ

  17. Sequestering CO2 in the Built Environment

    NASA Astrophysics Data System (ADS)

    Constantz, B. R.

    2009-12-01

    Calera’s Carbonate Mineralization by Aqueous Precipitation (CMAP) technology with beneficial reuse has been called, “game-changing” by Carl Pope, Director of the Sierra Club. Calera offers a solution to the scale of the carbon problem. By capturing carbon into the built environment through carbonate mineralization, Calera provides a sound and cost-effective alternative to Geologic Sequestration and Terrestrial Sequestration. The CMAP technology permanently converts carbon dioxide into a mineral form that can be stored above ground, or used as a building material. The process produces a suite of carbonate-containing minerals of various polymorphic forms. Calera product can be substituted into blends with ordinary Portland cements and used as aggregate to produce concrete with reduced carbon, carbon neutral, or carbon negative footprints. For each ton of product produced, approximately half a ton of carbon dioxide can be sequestered using the Calera process. Coal and natural gas are composed of predominately istopically light carbon, as the carbon in the fuel is plant-derived. Thus, power plant CO2 emissions have relatively low δ13C values.The carbon species throughout the CMAP process are identified through measuring the inorganic carbon content, δ13C values of the dissolved carbonate species, and the product carbonate minerals. Measuring δ13C allows for tracking the flue gas CO2 throughout the capture process. Initial analysis of the capture of propane flue gas (δ13C ˜ -25 ‰) with seawater (δ13C ˜ -10 ‰) and industrial brucite tailings from a retired magnesium oxide plant in Moss Landing, CA (δ13C ˜ -7 ‰ from residual calcite) produced carbonate mineral products with a δ13C value of ˜ -20 ‰. This isotopically light carbon, transformed from flue gas to stable carbonate minerals, can be transferred and tracked through the capture process, and finally to the built environment. CMAP provides an economical solution to global warming by producing

  18. Microbial Biosensors: Engineered Microorganisms as the Sensing Machinery

    PubMed Central

    Park, Miso; Tsai, Shen-Long; Chen, Wilfred

    2013-01-01

    Whole-cell biosensors are a good alternative to enzyme-based biosensors since they offer the benefits of low cost and improved stability. In recent years, live cells have been employed as biosensors for a wide range of targets. In this review, we will focus on the use of microorganisms that are genetically modified with the desirable outputs in order to improve the biosensor performance. Different methodologies based on genetic/protein engineering and synthetic biology to construct microorganisms with the required signal outputs, sensitivity, and selectivity will be discussed. PMID:23648649

  19. Non-Invasive Optical Biosensor for Probing Cell Signaling

    PubMed Central

    Fang, Ye

    2007-01-01

    Cell signaling mediated through a cellular target is encoded by spatial and temporal dynamics of downstream signaling networks. The coupling of temporal dynamics with spatial gradients of signaling activities guides cellular responses upon stimulation. Monitoring the integration of cell signaling in real time, if realized, would provide a new dimension for understanding cell biology and physiology. Optical biosensors including resonant waveguide grating (RWG) biosensor manifest a physiologically relevant and integrated cellular response related to dynamic redistribution of cellular matters, thus providing a non-invasive means for cell signaling study. This paper reviews recent progresses in biosensor instrumentation, and theoretical considerations and potential applications of optical biosensors for whole cell sensing.

  20. A Novel High-Throughput Cell-Based Assay Aimed at Identifying Inhibitors of DNA Metabolism in Bacteria

    PubMed Central

    Fan, Jun; de Jonge, Boudewijn L. M.; MacCormack, Kathy; Sriram, Shubha; McLaughlin, Robert E.; Plant, Helen; Preston, Marian; Fleming, Paul R.; Albert, Robert; Foulk, Melinda

    2014-01-01

    Bacterial biosensor strains can be useful tools for the discovery and characterization of antibacterial compounds. A plasmid-based reporter vector containing a transcriptional fusion between the recA promoter and green fluorescence protein gene was introduced into an Escherichia coli ΔtolC strain to create a biosensor strain that selectively senses inhibitors of DNA metabolism via the SOS response. The strain was used to develop a high-throughput assay to identify new inhibitors of DNA metabolism. Screening of the AstraZeneca compound library with this strain identified known inhibitors of DNA metabolism, as well as novel chemotypes. The cellular target of one novel series was elucidated as DNA gyrase through genetic characterization of laboratory-generated resistant mutants followed by 50% inhibitory concentration measurements in a DNA gyrase activity assay. These studies validated the use of this antibiotic biosensor strain to identify novel selective inhibitors of DNA metabolism by high-throughput screening. PMID:25246396

  1. Characterization of DNA-Hv1 histone interactions; discrimination of DNA size and shape.

    PubMed

    Papadakis, George; Tsortos, Achilleas; Mitsakakis, Konstantinos; Gizeli, Electra

    2010-03-01

    We have studied the formation of histone Hv1-DNA complexes using an acoustic biosensor and AFM imaging. Our results show that DNA and histone molecules aggregate into amorphous accumulations which form a compact rigid layer on the sensor's surface. By measuring changes in the acoustic wave amplitude, it was possible to titrate surface bound DNA with Hv1 and discriminate between DNA molecules of different size and shape. From the kinetic analysis of real time data, Keq was found equal to 3x10(5) M(-1). PMID:20085758

  2. 46 CFR 67.97 - United States built.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

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  3. 46 CFR 67.97 - United States built.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 2 2014-10-01 2014-10-01 false United States built. 67.97 Section 67.97 Shipping COAST... DOCUMENTATION OF VESSELS Build Requirements for Vessel Documentation § 67.97 United States built. To be considered built in the United States a vessel must meet both of the following criteria: (a) All...

  4. 46 CFR 67.97 - United States built.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 2 2013-10-01 2013-10-01 false United States built. 67.97 Section 67.97 Shipping COAST... DOCUMENTATION OF VESSELS Build Requirements for Vessel Documentation § 67.97 United States built. To be considered built in the United States a vessel must meet both of the following criteria: (a) All...

  5. 47 CFR 15.23 - Home-built devices.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

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  6. 47 CFR 15.23 - Home-built devices.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

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  7. 46 CFR 67.97 - United States built.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 2 2011-10-01 2011-10-01 false United States built. 67.97 Section 67.97 Shipping COAST... DOCUMENTATION OF VESSELS Build Requirements for Vessel Documentation § 67.97 United States built. To be considered built in the United States a vessel must meet both of the following criteria: (a) All...

  8. 47 CFR 15.23 - Home-built devices.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 1 2013-10-01 2013-10-01 false Home-built devices. 15.23 Section 15.23 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.23 Home-built... that the individual builder of home-built equipment may not possess the means to perform...

  9. 47 CFR 15.23 - Home-built devices.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 1 2014-10-01 2014-10-01 false Home-built devices. 15.23 Section 15.23 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.23 Home-built... that the individual builder of home-built equipment may not possess the means to perform...

  10. 47 CFR 15.23 - Home-built devices.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 47 Telecommunication 1 2012-10-01 2012-10-01 false Home-built devices. 15.23 Section 15.23 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.23 Home-built... that the individual builder of home-built equipment may not possess the means to perform...

  11. 46 CFR 67.97 - United States built.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 2 2010-10-01 2010-10-01 false United States built. 67.97 Section 67.97 Shipping COAST... DOCUMENTATION OF VESSELS Build Requirements for Vessel Documentation § 67.97 United States built. To be considered built in the United States a vessel must meet both of the following criteria: (a) All...

  12. Microbial fuel cells for biosensor applications.

    PubMed

    Yang, Huijia; Zhou, Minghua; Liu, Mengmeng; Yang, Weilu; Gu, Tingyue

    2015-12-01

    Microbial fuel cells (MFCs) face major hurdles for real-world applications as power generators with the exception of powering small sensor devices. Despite tremendous improvements made in the last two decades, MFCs are still too expensive to build and operate and their power output is still too small. In view of this, in recently years, intensive researches have been carried out to expand the applications into other areas such as acid and alkali production, bioremediation of aquatic sediments, desalination and biosensors. Unlike power applications, MFC sensors have the immediate prospect to be practical. This review covers the latest developments in various proposed biosensor applications using MFCs including monitoring microbial activity, testing biochemical oxygen demand, detection of toxicants and detection of microbial biofilms that cause biocorrosion. PMID:26272393

  13. Surface modified amorphous ribbon based magnetoimpedance biosensor.

    PubMed

    Kurlyandskaya, Galina V; Fal Miyar, Vanessa

    2007-04-15

    Magnetoimpedance (MI) changes due to surface modification of the sensitive element caused by human urine, were studied with the aim of creating a robust biosensor working on a principle of electrochemical magnetoimpedance spectroscopy. A biosensor prototype with an as-quenched amorphous ribbon sensitive element was designed and calibrated for a frequency range of 0.5-10 MHz at a current intensity of 60 mA. Measurements as a function of the exposure time were made both in a regime where chemical surface modification and MI measurements were separated as well as in a regime where they were done simultaneously. The MI variation was explained by the change of the surface magnetic anisotropy. It was shown that the magnetoimpedance effect can be successfully employed as a new option to probe the electric features of the Fe(5)Co(70)Si(15)B(10) amorphous ribbon magnetic electrode surface modified by human urine. PMID:16914305

  14. Bioconjugation and stabilisation of biomolecules in biosensors

    PubMed Central

    Drago, Guido A.

    2016-01-01

    Suitable bioconjugation strategies and stabilisation of biomolecules on electrodes is essential for the development of novel and commercially viable biosensors. In the present review, the functional groups that comprise the selectable targets for practical bioconjugation methods are discussed. We focus on describing the most common immobilisation techniques used in biosensor construction, which are classified into irreversible and reversible methods. Concerning the stability of proteins, the two main types of stability may be defined as (i) storage or shelf stability, and (ii) operational stability. Both types of stability are explained, as well as the introduction of an electrophoretic technique for predicting protein–polymer interactions. In addition, solution and dry stabilisation as well as stabilisation using the covalent immobilisation of proteins are discussed including possible factors that influence stability. Finally, the integration of nanomaterials, such as magnetic particles, with protein immobilisation is discussed in relation to protein stability studies. PMID:27365036

  15. Bioconjugation and stabilisation of biomolecules in biosensors.

    PubMed

    Liébana, Susana; Drago, Guido A

    2016-06-30

    Suitable bioconjugation strategies and stabilisation of biomolecules on electrodes is essential for the development of novel and commercially viable biosensors. In the present review, the functional groups that comprise the selectable targets for practical bioconjugation methods are discussed. We focus on describing the most common immobilisation techniques used in biosensor construction, which are classified into irreversible and reversible methods. Concerning the stability of proteins, the two main types of stability may be defined as (i) storage or shelf stability, and (ii) operational stability. Both types of stability are explained, as well as the introduction of an electrophoretic technique for predicting protein-polymer interactions. In addition, solution and dry stabilisation as well as stabilisation using the covalent immobilisation of proteins are discussed including possible factors that influence stability. Finally, the integration of nanomaterials, such as magnetic particles, with protein immobilisation is discussed in relation to protein stability studies. PMID:27365036

  16. Modelling Carbon Nanotubes-Based Mediatorless Biosensor

    PubMed Central

    Baronas, Romas; Kulys, Juozas; Petrauskas, Karolis; Razumiene, Julija

    2012-01-01

    This paper presents a mathematical model of carbon nanotubes-based mediatorless biosensor. The developed model is based on nonlinear non-stationary reaction-diffusion equations. The model involves four layers (compartments): a layer of enzyme solution entrapped on a terylene membrane, a layer of the single walled carbon nanotubes deposited on a perforated membrane, and an outer diffusion layer. The biosensor response and sensitivity are investigated by changing the model parameters with a special emphasis on the mediatorless transfer of the electrons in the layer of the enzyme-loaded carbon nanotubes. The numerical simulation at transient and steady state conditions was carried out using the finite difference technique. The mathematical model and the numerical solution were validated by experimental data. The obtained agreement between the simulation results and the experimental data was admissible at different concentrations of the substrate. PMID:23012537

  17. Modelling carbon nanotubes-based mediatorless biosensor.

    PubMed

    Baronas, Romas; Kulys, Juozas; Petrauskas, Karolis; Razumiene, Julija

    2012-01-01

    This paper presents a mathematical model of carbon nanotubes-based mediatorless biosensor. The developed model is based on nonlinear non-stationary reaction-diffusion equations. The model involves four layers (compartments): a layer of enzyme solution entrapped on a terylene membrane, a layer of the single walled carbon nanotubes deposited on a perforated membrane, and an outer diffusion layer. The biosensor response and sensitivity are investigated by changing the model parameters with a special emphasis on the mediatorless transfer of the electrons in the layer of the enzyme-loaded carbon nanotubes. The numerical simulation at transient and steady state conditions was carried out using the finite difference technique. The mathematical model and the numerical solution were validated by experimental data. The obtained agreement between the simulation results and the experimental data was admissible at different concentrations of the substrate. PMID:23012537

  18. Recent advances in biosensor based endotoxin detection.

    PubMed

    Das, A P; Kumar, P S; Swain, S

    2014-01-15

    Endotoxins also referred to as pyrogens are chemically lipopolysaccharides habitually found in food, environment and clinical products of bacterial origin and are unavoidable ubiquitous microbiological contaminants. Pernicious issues of its contamination result in high mortality and severe morbidities. Standard traditional techniques are slow and cumbersome, highlighting the pressing need for evoking agile endotoxin detection system. The early and prompt detection of endotoxin assumes prime importance in health care, pharmacological and biomedical sectors. The unparalleled recognition abilities of LAL biosensors perched with remarkable sensitivity, high stability and reproducibility have bestowed it with persistent reliability and their possible fabrication for commercial applicability. This review paper entails an overview of various trends in current techniques available and other possible alternatives in biosensor based endotoxin detection together with its classification, epidemiological aspects, thrust areas demanding endotoxin control, commercially available detection sensors and a revolutionary unprecedented approach narrating the influence of omics for endotoxin detection. PMID:23934306

  19. Biosensors for waterborne viruses: Detection and removal.

    PubMed

    Altintas, Zeynep; Gittens, Micah; Pocock, Jack; Tothill, Ibtisam E

    2015-08-01

    Detection of waterborne viruses is important to eliminate and control their harmful effect as pathogens. Hence, the use of rapid and sensitive detection technologies is critically important as they can aid in investigating outbreaks and help in developing prevention strategies. To date range of viruses can contaminate drinking water sources, causing illnesses such as diarrhoea, pneumonia and gastroenteritis which can result in death. Due to their small size (nm) their complete removal from water can be difficult with current water treatment processes while being resistant to disinfectants. Available techniques for virus detection include filtration technologies, enzyme-linked immunosorbent assays and polymerase chain reaction. Although each technique has limitations, the use of biosensor technology with smart affinity materials and nanomaterials can show great potential in sensing viruses in water samples. This review reports on the latest technologies used for waterborne virus removal and detection with focus on rapid detection using biosensors. PMID:26005094

  20. A portable array biosensor for food safety

    NASA Astrophysics Data System (ADS)

    Golden, Joel P.; Ngundi, Miriam M.; Shriver-Lake, Lisa C.; Taitt, Chris R.; Ligler, Frances S.

    2004-11-01

    An array biosensor developed for simultaneous analysis of multiple samples has been utilized to develop assays for toxins and pathogens in a variety of foods. The biochemical component of the multi-analyte biosensor consists of a patterned array of biological recognition elements immobilized on the surface of a planar waveguide. A fluorescence assay is performed on the patterned surface, yielding an array of fluorescent spots, the locations of which are used to identify what analyte is present. Signal transduction is accomplished by means of a diode laser for fluorescence excitation, optical filters and a CCD camera for image capture. A laptop computer controls the miniaturized fluidics system and image capture. Results for four mycotoxin competition assays in buffer and food samples are presented.

  1. Amperometric biosensors based on carbon composite transducers

    NASA Astrophysics Data System (ADS)

    Lu, Fang

    1998-12-01

    Much current work in analytical chemistry is devoted to design of biosensors. One particular area in this field is the development of enzyme-based amperometric biosensors for the quantitative determination of a series of substrates in clinical, environmental, industrial and agricultural significance. This dissertation focuses on the design of improved amperometric biosensors based on carbon composite transducers. The use of metallized carbons as transducer materials results in remarkably selective amperometric biosensors. Such enzyme-based transducers eliminate major electroactive interferences, and hence circumvent the need for mediators or membrane barriers. The remarkable selectivity of metal-dispersed carbons is attributed to their strong, preferential, electrocatalytic capacity towards the reductive detection of biologically-generated hydrogen peroxide. Such electrocatalytic activity allows metal-dispersed biosensors to be operated at the optimal potential region between +0.1 and -0.2 V, where the unwanted reactions are neglected resulting in the lowest noise level. Several new materials (e.g., ruthenium on carbon, rhodium on carbon, etc.) and constructions (e.g., carbon fiber, electrochemical co-deposition transducer, etc.) were applied in the development of novel enzyme-based transducers in order to improve the selectivity and applicability of amperometric biosensors. The susceptibility of first-generation oxidase amperometric biosensing to oxygen fluctuations can be improved by using oxygen-rich fluorocarbons as the pasting binders in carbon paste enzyme transducers. Such binders provide an internal supply of oxygen resulting in efficient detection in oxygen-deficit conditions. In particular, the use of poly-chlorotrifluorethylene (Kel-F) oil as carbon paste binder results in a well-defined response and an identical signal up to 40 mM glucose in both the presence and absence of oxygen. Comparing with mediated or wired enzyme-based transducers, such internal

  2. Long Wavelength Fluorescence Ratiometric Zinc Biosensor

    PubMed Central

    Zeng, Hui Hui; Matveeva, Evgenia; Stoddard, Andrea K.; Fierke, Carol A.; Thompson, Richard B.

    2013-01-01

    A protein-based emission ratiometric fluorescence biosensor is described that exhibits sensitivity to free zinc ion solutions down to picomolar concentrations. Ratiometric measurements are widely used to assure accurate quantitation, and emission ratios are preferred for laser scanning microscopes such as confocal fluorescence microscopes. The relatively long emission wavelengths used are well suited to studies in tissues and other matrices which exhibit significant fluorescence background, and the apo-carbonic anhydrase moiety recognizes zinc ion with high and controllable specificity. PMID:23345045

  3. Biosensors Related LLT: Physiopathology, Background and Rationale

    NASA Astrophysics Data System (ADS)

    Palmieri, Beniamino; Capone, Stefania

    2011-08-01

    The Authors describe a new biosensor-related approach to LLT delivery, tailored to homogeneously integrate the external energy to the maximum patient compliance and receptivity; this project is based on the work hypothesis, by a pilot clinical trial and experimental animals (liver crush injury in the rat) confirmed that the diodes number, frequency, intensity, and pulses when synchronized with the vital parameters induce maximal physiological response in terms of regeneration and recovery.

  4. Dual-mode acoustic wave biosensors microarrays

    NASA Astrophysics Data System (ADS)

    Auner, Gregory W.; Shreve, Gina; Ying, Hao; Newaz, Golam; Hughes, Chantelle; Xu, Jianzeng

    2003-04-01

    We have develop highly sensitive and selective acoustic wave biosensor arrays with signal analysis systems to provide a fingerprint for the real-time identification and quantification of a wide array of bacterial pathogens and environmental health hazards. We have developed an unique highly sensitive dual mode acoustic wave platform prototype that, when combined with phage based selective detection elements, form a durable bacteria sensor. Arrays of these new real-time biosensors are integrated to form a biosensor array on a chip. This research and development program optimizes advanced piezoelectric aluminum nitride wide bandgap semiconductors, novel micromachining processes, advanced device structures, selective phage displays development and immobilization techniques, and system integration and signal analysis technology to develop the biosensor arrays. The dual sensor platform can be programmed to sense in a gas, vapor or liquid environment by switching between acoustic wave resonate modes. Such a dual mode sensor has tremendous implications for applications involving monitoring of pathogenic microorganisms in the clinical setting due to their ability to detect airborne pathogens. This provides a number of applications including hospital settings such as intensive care or other in-patient wards for the reduction of nosocomial infections and maintenance of sterile environments in surgical suites. Monitoring for airborn pathogen transmission in public transportation areas such as airplanes may be useful for implementation of strategies for redution of airborn transmission routes. The ability to use the same sensor in the liquid sensing mode is important for tracing the source of airborn pathogens to local liquid sources. Sensing of pathogens in saliva will be useful for sensing oral pathogens and support of decision-making strategies regarding prevention of transmission and support of treatment strategies.

  5. Integrated-optical directional coupler biosensor

    NASA Astrophysics Data System (ADS)

    Luff, B. J.; Harris, R. D.; Wilkinson, J. S.; Wilson, R.; Schiffrin, D. J.

    1996-04-01

    We present measurements of biomolecular binding reactions, using a new type of integrated-optical biosensor based on a planar directional coupler structure. The device is fabricated by Ag+ - Na+ ion exchange in glass, and definition of the sensing region is achieved by use of transparent fluoropolymer isolation layers formed by thermal evaporation. The suitability of the sensor for application to the detection of environmental pollutants is considered.

  6. Biosensor discovery of thyroxine transport disrupting chemicals

    SciTech Connect

    Marchesini, Gerardo R. Meimaridou, Anastasia; Haasnoot, Willem; Meulenberg, Eline; Albertus, Faywell; Mizuguchi, Mineyuki; Takeuchi, Makoto; Irth, Hubertus; Murk, Albertinka J.

    2008-10-01

    Ubiquitous chemicals may interfere with the thyroid system that is essential in the development and physiology of vertebrates. We applied a surface plasmon resonance (SPR) biosensor-based screening method for the fast screening of chemicals with thyroxine (T4) transport disrupting activity. Two inhibition assays using the main thyroid hormone transport proteins, T4 binding globulin (TBG) and transthyretin (TTR), in combination with a T4-coated biosensor chip were optimized and automated for screening chemical libraries. The transport protein-based biosensor assays were rapid, high throughput and bioeffect-related. A library of 62 chemicals including the natural hormones, polychlorinated biphenyls (PCBs), polybrominated diphenylethers (PBDEs) and metabolites, halogenated bisphenol A (BPA), halogenated phenols, pharmaceuticals, pesticides and other potential environmentally relevant chemicals was tested with the two assays. We discovered ten new active compounds with moderate to high affinity for TBG with the TBG assay. Strikingly, the most potent binding was observed with hydroxylated metabolites of the brominated diphenyl ethers (BDEs) BDE 47, BDE 49 and BDE 99, that are commonly found in human plasma. The TTR assay confirmed the activity of previously identified hydroxylated metabolites of PCBs and PBDEs, halogenated BPA and genistein. These results show that the hydroxylated metabolites of the ubiquitous PBDEs not only target the T4 transport at the TTR level, but also, and to a great extent, at the TBG level where most of the T4 in humans is circulating. The optimized SPR biosensor-based transport protein assay is a suitable method for high throughput screening of large libraries for potential thyroid hormone disrupting compounds.

  7. Scalable Production of Molybdenum Disulfide Based Biosensors.

    PubMed

    Naylor, Carl H; Kybert, Nicholas J; Schneier, Camilla; Xi, Jin; Romero, Gabriela; Saven, Jeffery G; Liu, Renyu; Johnson, A T Charlie

    2016-06-28

    We demonstrate arrays of opioid biosensors based on chemical vapor deposition grown molybdenum disulfide (MoS2) field effect transistors (FETs) coupled to a computationally redesigned, water-soluble variant of the μ-opioid receptor (MOR). By transferring dense films of monolayer MoS2 crystals onto prefabricated electrode arrays, we obtain high-quality FETs with clean surfaces that allow for reproducible protein attachment. The fabrication yield of MoS2 FETs and biosensors exceeds 95%, with an average mobility of 2.0 cm(2) V(-1) s(-1) (36 cm(2) V(-1) s(-1)) at room temperature under ambient (in vacuo). An atomic length nickel-mediated linker chemistry enables target binding events that occur very close to the MoS2 surface to maximize sensitivity. The biosensor response calibration curve for a synthetic opioid peptide known to bind to the wild-type MOR indicates binding affinity that matches values determined using traditional techniques and a limit of detection ∼3 nM (1.5 ng/mL). The combination of scalable array fabrication and rapid, precise binding readout enabled by the MoS2 transistor offers the prospect of a solid-state drug testing platform for rapid readout of the interactions between novel drugs and their intended protein targets. PMID:27227361

  8. Raman Spectroscopy Cell-based Biosensors

    PubMed Central

    Notingher, Ioan

    2007-01-01

    One of the main challenges faced by biodetection systems is the ability to detect and identify a large range of toxins at low concentrations and in short times. Cell-based biosensors rely on detecting changes in cell behaviour, metabolism, or induction of cell death following exposure of live cells to toxic agents. Raman spectroscopy is a powerful technique for studying cellular biochemistry. Different toxic chemicals have different effects on living cells and induce different time-dependent biochemical changes related to cell death mechanisms. Cellular changes start with membrane receptor signalling leading to cytoplasmic shrinkage and nuclear fragmentation. The potential advantage of Raman spectroscopy cell-based systems is that they are not engineered to respond specifically to a single toxic agent but are free to react to many biologically active compounds. Raman spectroscopy biosensors can also provide additional information from the time-dependent changes of cellular biochemistry. Since no cell labelling or staining is required, the specific time dependent biochemical changes in the living cells can be used for the identification and quantification of the toxic agents. Thus, detection of biochemical changes of cells by Raman spectroscopy could overcome the limitations of other biosensor techniques, with respect to detection and discrimination of a large range of toxic agents. Further developments of this technique may also include integration of cellular microarrays for high throughput in vitro toxicological testing of pharmaceuticals and in situ monitoring of the growth of engineered tissues.

  9. Hybrid nano plasmonics for integrated biosensor

    NASA Astrophysics Data System (ADS)

    Lin, Chii-Wann; Lee, Jun-Haw; Chiu, Nan-Fu; Lee, Szu-Yuan; Liu, Kou-Chen; Tsai, Feng-Yu; Yen, Chia-Yu; Lee, Chun-Nan

    2009-11-01

    SPR biosensor with OLED and nano-grating for HBV LAMP product detection is reported. Directional emissions by grating-coupler match the resonant condition of SP modes. Concentration changes result in color shift at specific angle. Real time detection of virus load down to 5 copies/25 ul can be achieved in 30 minutes. Surface plasmon Resonant (SPR) biosensor has been used for quantitative measurement of molecular interactions for its advantages of high sensitivity, label-free and real-time detection. In this paper, we report recent efforts on further enhancement of SPR biosensors by the heterogeneous integration of organic electroluminescence light source and nano-grating structure for the feasibility study on the fast and high sensitivity detection of HBV isothermal amplification products, Mg2P2O7. We demonstrated the surface plasmon coupled through hybrid nano-grating structure has highly directional emissions corresponding to the resonant condition of surface plasmon modes on the Au/air interface and controllable plasmonics band-gap by pitch modulation. SPGCE resulted in color change from yellowish green to orange at a certain viewing angle, when contacting glucose with concentration increasing from 10 to 40%.

  10. Design of nanostructured-based glucose biosensors

    NASA Astrophysics Data System (ADS)

    Komirisetty, Archana; Williams, Frances; Pradhan, Aswini; Konda, Rajini B.; Dondapati, Hareesh; Samantaray, Diptirani

    2012-04-01

    This paper presents the design of glucose sensors that will be integrated with advanced nano-materials, bio-coatings and electronics to create novel devices that are highly sensitive, inexpensive, accurate, and reliable. In the work presented, a glucose biosensor and its fabrication process flow have been designed. The device is based on electrochemical sensing using a working electrode with bio-functionalized zinc oxide (ZnO) nano-rods. Among all metal oxide nanostructures, ZnO nano-materials play a significant role as a sensing element in biosensors due to their properties such as high isoelectric point (IEP), fast electron transfer, non-toxicity, biocompatibility, and chemical stability which are very crucial parameters to achieve high sensitivity. Amperometric enzyme electrodes based on glucose oxidase (GOx) are used due to their stability and high selectivity to glucose. The device also consists of silicon dioxide and titanium layers as well as platinum working and counter electrodes and a silver/silver chloride reference electrode. Currently, the biosensors are being fabricated using the process flow developed. Once completed, the sensors will be bio-functionalized and tested to characterize their performance, including their sensitivity and stability.

  11. Cultured neuronal networks as environmental biosensors.

    PubMed

    O'Shaughnessy, Thomas J; Gray, Samuel A; Pancrazio, Joseph J

    2004-01-01

    Contamination of water by toxins, either intentionally or unintentionally, is a growing concern for both military and civilian agencies and thus there is a need for systems capable of monitoring a wide range of natural and industrial toxicants. The EILATox-Oregon Workshop held in September 2002 provided an opportunity to test the capabilities of a prototype neuronal network-based biosensor with unknown contaminants in water samples. The biosensor is a portable device capable of recording the action potential activity from a network of mammalian neurons grown on glass microelectrode arrays. Changes in the action potential fi ring rate across the network are monitored to determine exposure to toxicants. A series of three neuronal networks derived from mice was used to test seven unknown samples. Two of these unknowns later were revealed to be blanks, to which the neuronal networks did not respond. Of the five remaining unknowns, a significant change in network activity was detected for four of the compounds at concentrations below a lethal level for humans: mercuric chloride, sodium arsenite, phosdrin and chlordimeform. These compounds--two heavy metals, an organophosphate and an insecticide--demonstrate the breadth of detection possible with neuronal networks. The results generated at the workshop show the promise of the neuronal network biosensor as an environmental detector but there is still considerable effort needed to produce a device suitable for routine environmental threat monitoring. PMID:15478174

  12. Sensitive-cell-based fish chromatophore biosensor

    NASA Astrophysics Data System (ADS)

    Plant, Thomas K.; Chaplen, Frank W.; Jovanovic, Goran; Kolodziej, Wojtek; Trempy, Janine E.; Willard, Corwin; Liburdy, James A.; Pence, Deborah V.; Paul, Brian K.

    2004-07-01

    A sensitive biosensor (cytosensor) has been developed based on color changes in the toxin-sensitive colored living cells of fish. These chromatophores are highly sensitive to the presence of many known and unknown toxins produced by microbial pathogens and undergo visible color changes in a dose-dependent manner. The chromatophores are immobilized and maintained in a viable state while potential pathogens multiply and fish cell-microbe interactions are monitored. Low power LED lighting is used to illuminate the chromatophores which are magnified using standard optical lenses and imaged onto a CCD array. Reaction to toxins is detected by observing changes is the total area of color in the cells. These fish chromatophores are quite sensitive to cholera toxin, Staphococcus alpha toxin, and Bordatella pertussis toxin. Numerous other toxic chemical and biological agents besides bacterial toxins also cause readily detectable color effects in chromatophores. The ability of the chromatophore cell-based biosensor to distinguish between different bacterial pathogens was examined. Toxin producing strains of Salmonella enteritis, Vibrio parahaemolyticus, and Bacillus cereus induced movement of pigmented organelles in the chromatophore cells and this movement was measured by changes in the optical density over time. Each bacterial pathogen elicited this measurable response in a distinctive and signature fashion. These results suggest a chromatophore cell-based biosensor assay may be applicable for the detection and identification of virulence activities associated with certain air-, food-, and water-borne bacterial pathogens.

  13. Biosensor for underwater chemical sensing (Invited Paper)

    NASA Astrophysics Data System (ADS)

    Kusterbeck, Anne W.; Deschamps, Jeffrey R.; Charles, Paul T.

    2005-05-01

    Emerging biosensor approaches may prove useful in reducing false positives and improving detection probabilities for unexploded ordnance (UXO) and underwater explosives. NRL researchers previously developed a biosensor that was field-tested and validated for use in environmental remediation to detect explosives in groundwater. The sensor relies on the selective recognition by antibodies of target analytes, including the common explosives TNT and RDX. Laboratory work has demonstrated that sensors based on these displacement immunoassay formats can detect explosives at the part-per-trillion level in seawater. More recently, participating in an Office of Naval Research program on Chemical Sensing in the Marine Environment (CSME), tests were conducted in controlled underwater experiments at San Clemente, CA and Duck, NC. Simulated UXO targets, autonomous underwater vehicles (AUV) and multiple sensor approaches were used to demonstrate the feasibility of underwater chemical sensing. Efforts are now underway to integrate the biosensor into an underwater platform as part of a broader sensor system. We will describe results of these studies and outline possible operational scenarios for applications in harbor security.

  14. Biosensor UUV payload for underwater detection

    NASA Astrophysics Data System (ADS)

    Kusterbeck, Anne W.; Charles, Paul T.; Melde, Brian J.; Trammell, Scott A.; Adams, André A.; Deschamps, Jeffrey R.

    2010-04-01

    Increased emphasis on maritime domain awareness and port security has led to the development of unmanned underwater vehicles (UUVs) capable of extended missions. These systems rely most frequently on well-developed side scan sonar and acoustic methods to locate potential targets. The Naval Research Laboratory (NRL) is developing biosensors for underwater explosives detection that complement acoustic sensors and can be used as UUV payloads to monitor areas for port and harbor security or in detection of underwater unexploded ordnance (UXO) and biochemical threats. The prototype sensor has recently been demonstrated to detect explosives in seawater at trace levels when run in a continuous sampling mode. To overcome ongoing issues with sample preparation and facilitate rapid detection at trace levels in a marine environment, we have been developing new mesoporous materials for in-line preconcentration of explosives and other small molecules, engineering microfluidic components to improve the signal, and testing alternative signal transduction methods. Additional work is being done to optimize the optical components and sensor response time. Highlights of these current studies and our ongoing efforts to integrate the biosensor with existing detection technologies to reduce false positives are described. In addition, we present the results of field tests that demonstrate the prototype biosensor performance as a UUV payload.

  15. Electrowetting on dielectric digital microfluidic platform with nanostructured biosensor interface for enhanced two-dimensional surface plasmon resonance imaging detection

    NASA Astrophysics Data System (ADS)

    Malic, Lidija

    The sensitive and specific detection of biomolecular interactions is at the heart of many routine analyses in fundamental research, medical diagnosis and environmental monitoring. In contrast to laborious and costly multiwell plate assays, recent years have witnessed a significant progress in miniaturized and integrated biosensors, such as surface plasmon resonance (SPR), tailored to these applications. While the design of various SPR biosensors has been described in literature, a robust, multichannel, low-cost and highly sensitive solution has not yet been presented. Specifically, an integrated system that can allow surface functionalization in array format, low-volume multichannel fluidic interfacing, and increased sensitivity is sought. This thesis describes a novel electro-wetting-on-dielectric (EWOD) digital microfluidic device with integrated nanostructured biosensor interface that addresses the aforementioned issues for enhanced surface plasmon resonance imaging (SPRi) detection. We have taken the opportunity of the most recent advances in microfabrication, nanotechnology and SPR technique to develop this integrated platform. EWOD device is employed for the dynamic immobilization of bioreceptors on SPRi biosensor surface in an array fashion from sub-muL volume solutions. Programmable EWOD electric interface allows the application of an electric field at the biosensor surface for active control of the immobilized probe density and orientation, enhancing SPRi detection. Two-dimensional SPRi detection is achieved by coupling the EWOD device to SPRi instrumentation. Parallel manipulation of individual droplets allows more efficient exploitation of the biosensor surface by separating different samples for simultaneous and selective SPRi detection. Periodic gold structures (nanoposts, nanogratings and nanogrooves) residing on a surface of glass and plastic substrates are investigated to improve the SPRi sensitivity. The corresponding electromagnetic field

  16. Amorphous/nanocrystalline silicon biosensor for the specific identification of unamplified nucleic acid sequences using gold nanoparticle probes

    NASA Astrophysics Data System (ADS)

    Martins, Rodrigo; Baptista, Pedro; Raniero, Leandro; Doria, Gonçalo; Silva, Leonardo; Franco, Ricardo; Fortunato, Elvira

    2007-01-01

    Amorphous/nanocrystalline silicon pi 'ii'n devices fabricated on micromachined glass substrates are integrated with oligonucleotide-derivatized gold nanoparticles for a colorimetric detection method. The method enables the specific detection and quantification of unamplified nucleic acid sequences (DNA and RNA) without the need to functionalize the glass surface, allowing for resolution of single nucleotide differences between DNA and RNA sequences—single nucleotide polymorphism and mutation detection. The detector's substrate is glass and the sample is directly applied on the back side of the biosensor, ensuring a direct optical coupling of the assays with a concomitant maximum photon capture and the possibility to reuse the sensor.

  17. In vitro evaluation of fluorescence glucose biosensor response.

    PubMed

    Aloraefy, Mamdouh; Pfefer, T Joshua; Ramella-Roman, Jessica C; Sapsford, Kim E

    2014-01-01

    Rapid, accurate, and minimally-invasive glucose biosensors based on Förster Resonance Energy Transfer (FRET) for glucose measurement have the potential to enhance diabetes control. However, a standard set of in vitro approaches for evaluating optical glucose biosensor response under controlled conditions would facilitate technological innovation and clinical translation. Towards this end, we have identified key characteristics and response test methods, fabricated FRET-based glucose biosensors, and characterized biosensor performance using these test methods. The biosensors were based on competitive binding between dextran and glucose to concanavalin A and incorporated long-wavelength fluorescence dye pairs. Testing characteristics included spectral response, linearity, sensitivity, limit of detection, kinetic response, reversibility, stability, precision, and accuracy. The biosensor demonstrated a fluorescence change of 45% in the presence of 400 mg/dL glucose, a mean absolute relative difference of less than 11%, a limit of detection of 25 mg/dL, a response time of 15 min, and a decay in fluorescence intensity of 72% over 30 days. The battery of tests presented here for objective, quantitative in vitro evaluation of FRET glucose biosensors performance have the potential to form the basis of future consensus standards. By implementing these test methods for a long-visible-wavelength biosensor, we were able to demonstrate strengths and weaknesses with a new level of thoroughness and rigor. PMID:25006996

  18. A versatile biosensor device for continuous biomedical monitoring.

    PubMed

    Rhemrev-Boom, M M; Korf, J; Venema, K; Urban, G; Vadgama, P

    2001-12-01

    Although biosensors are by means suitable for continuous biomedical monitoring, due to fouling and blood clotting, in vivo performance is far from optimal. For this reason, ultrafiltration, microdialysis or open tubular flow is frequently used as interface. To secure quantitative recoveries of the analyte of interest, sampling at submicrolitre level will be necessary which in turn necessitates the development of small and versatile biosensor devices. Here, a miniaturised biosensor device, which directly can be connected to various interfaces will be presented. The biosensor device consists of a pulsefree pump and a biosensor with an internal volume of 10-20 nl. In this article, the production as well as the construction of the flow-through cell of the biosensor will be discussed. The advantages and disadvantages of several production processes will be demonstrated and a detailed protocol for the production of such a nanoliter flow-through cell will be presented. With respect to the bio-selector, several permselective membranes have been tested on their performance characteristics. Results obtained with these biosensors will be presented and discussed. Finally, a protocol based upon in situ electropolymerisation for the immobilisation of the biological component was defined and several biosensors based upon this principle have been produced and tested for the monitoring of glucose respectively lactate. To demonstrate, data obtained during a variety of in vivo studies at different clinical relevant applications will be presented. PMID:11679262

  19. Homemade Bienzymatic-Amperometric Biosensor for Beverages Analysis

    ERIC Educational Resources Information Center

    Blanco-Lopez, M. C.; Lobo-Castanon, M. J.; Miranda-Ordieres, A. J.

    2007-01-01

    The construction of an amperometric biosensor for glucose analysis is described demonstrating that the analysis is easy to perform and the biosensor gives good analytical performance. This experiment helped the students to acquire problem-solving and teamwork skills, allowing them to reach a high level of independent and critical thought.

  20. Multiple Pathogen Detection Using Biosensors: Advancements and Challenges

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Advancements in biosensor research have considerably impacted clinical diagnostics for human health. Efforts in capitalizing on the sensitivity of biosensors for food pathogen detection are evident in the food safety/security research community. For practical application with foods that normally h...