The Escherichia coli O157:H7 DNA detection on a gold nanoparticle-enhanced piezoelectric biosensor

This paper presents development of a quartz crystal microbalance （QCM） biosensor for real-time detection of E. coil O157：H7 DNA based on nanogold particles amplification. Many inner Au nanoparticles were immobilized onto the thioled surface of the Au electrode, then more specific thiolated sin- gle-stranded DNA （ssDNA） probes could be fixed through Au-SH bonding. The hybridization was induced by exposing the ssDNA probe to the complementary target DNA of E. coli O157：H7 gene eaeA, then resulted in a mass change and corresponding frequency shifts （ △f ） of the QCM. The outer avidin-coated Au nanoparticles could combine with the target DNA to increase the mass. The electrochemical techniques, cyclic voltammetry （CV） and electrochemical impedance spectroscopy （EIS） were adopted to manifest and character each step. The target DNA corresponding to 2.0×10^3 colony forming unit （CFU）/mL E. coil O157：H7 cells can be detected by this biosensor, so it is practical to develop a sensitive and effective QCM biosensor for pathogenic bacteria detection based on specific DNA analysis. The piezoelectric biosensing system has potential for further applications, such as food safety and environment monitoring, and this approach lays the groundwork for incorporating the method into an integrated system for in-field bacteria detection.     

The Escherichia coli O157:H7 DNA detection on a gold nanoparticle-enhanced piezoelectric biosensor

This paper presents development of a quartz crystal microbalance (QCM) biosensor for real-time de- tection of E. coli O157:H7 DNA based on nanogold particles amplification. Many inner Au nanoparticles were immobilized onto the thioled surface of the Au electrode, then more specific thiolated sin- gle-stranded DNA (ssDNA) probes could be fixed through Au-SH bonding. The hybridization was in- duced by exposing the ssDNA probe to the complementary target DNA of E. coli O157:H7 gene eaeA, then resulted in a mass change and corresponding frequency shifts ( △f ) of the QCM. The outer avidin-coated Au nanoparticles could combine with the target DNA to increase the mass. The electro- chemical techniques, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were adopted to manifest and character each step. The target DNA corresponding to 2.0×103 colony forming unit (CFU)/mL E. coli O157:H7 cells can be detected by this biosensor, so it is practical to develop a sensitive and effective QCM biosensor for pathogenic bacteria detection based on specific DNA analy- sis. The piezoelectric biosensing system has potential for further applications, such as food safety and environment monitoring, and this approach lays the groundwork for incorporating the method into an integrated system for in-field bacteria detection.     

Determination of the total concentration of casein and β-lactoglobulin by indirect competitive ELISA in milk and dairy products

The main purpose of this work was to develop an assay as a means of quality control in milk and dairy products. As specific and high abundant proteins in milk and diary products, casein and β-lactoglobulin were chosen as the targets, and an indirect competitive enzyme-linked immunosorbent assay （ELISA） was developed for determination of the total concentration of casein and β-lactoglobulin. The detection limit of the assay was 79.07 ng/mL, and a working range of the calibration curve was from 140.6 to 36,000 ng/mL. The intrassay and inter-assay coefficients of variations were 4.7 % and 6.5 %, respectively. The recoveries ranged from 94.1% to 106.6 %. The developed ELISA assay was used to detect the total con- centration of casein and β-lactoglobulin in liquid milk and milk powder. The results indicated that the developed assay was very specific, and the addition of nonprotein nitrogen （such as melamine, cyanuric acid, and urea） and vegetable protein did not affect the detection. The good performance of the assay makes it suitable for use in the quality control of milk and dairy products.     

A high-sensitive detection method for carvone odor by implanted electrodes in rat olfactory bulb

Mammalian olfactory systems have extraordi- nary ability to sense and identify various trace odorants. Taking advantages of cell culture and micro-fabrication technologies, olfactory cell- or tissue-based biosensor represent a promising platform for in vitro odorant detec- tion. However, in vitro conditions lead to shortened cell/ tissue survivals, and the working life of neuron chips is short. The purpose of this study is to develop an in vivo recording and analyzing method for long-term and repeatable detection of odor stimulation. In this study, we implanted penetrating micro-wire array electrode into the olfactory bulb of conscious rats to obtain odor-evoked electrophysiological activities. Then, we investigated the response of ensembles of mitral/tufted cells to stimulation with carvone at a number of concentrations in time and frequency domains. The stable, repeatable odorant responses from up to 16 neural regions could be obtained for at least 3 weeks. Further, we explored the concentration detection sensitivity limitation of developed method, and found the detection low limit of carvone was below 10-a~ mol/L. The result demonstrates that the concentra- tion range of in vivo odorant detection method is much wider than in vitro method.     

Subtyping of type A influenza by sequencing the variable regions of HA gene specifically amplified with RT-PCR

The outbreak of a novel influenza A （H1N1） virus across the globe poses a threat to human health. It is of paramount importance to develop a rapid, reliable and inexpensive diagnostic procedure. Based on the bioinformatic information from public database, primers specific for influenza A virus surface protein haemagglutinin （HA） of several subtypes （including H1, H2, H3, H5, H7 and H9） were designed. Primer-specific PCR products were subiected to sequencing for accurately distinguishing H1 and H3 subtypes from others. This sequencing-based detection method will not only be applied to rapid detection and simultaneous subtype identification of new influenza A virus H1N1, but also provide the strategies to monitor other new types of influenza virus with explosive potential.     

Novel protein detection method based on proximity-dependent polymerase reaction and aptamers

In recent years, specific detection of proteins is one of the hot issues about aptamers in proteomics. Here we reported a simple, sensitive and specific proximity-dependent protein assay with dual DNA aptamers. Thrombin was used as the model protein, and two aptamer probes with complementary sequence at 3′-end were designed for the two distinct epitopes of the protein. Association of the two aptamers with thrombin resulted in stable hybrids due to the proximity of 3′-end, then polymerase reaction was induced. The amount of obtained dsDNA was indicated using the fluorescence dye Sybr Green I. The results showed that the initial velocity of polymerase reaction had a positive correlation with concentration of thrombin. The advantages of this dual-aptamer-based approach included simple and flexible design of aptamer probes, high selectivity and high sensitivity. The detection limit was 6.9 pmol/L.     

Optical detection of acetylcholine esterase based on CdTe quantum dots

In this study, we have constructed a simple, sensitive and rapid biosensor for detection of acetylcholine esterase (AChE) based on CdTe quantum dots (QDs). The detection limit of AChE by one-step enzyme reaction based on the thiolglycolic acid (TGA) stabilized QDs (TGA-QDs) was 10 U/L and the linear range was 10-100 and 100-1200 U/L, respectively. The detection limit of AChE by two-step enzyme reaction based on the 3-mercaptopropionic acid (MGA) stabilized QDs (MGA-QDs) was found to be 20 U/L and the linear range was 100-2500 U/L. The experimental conditions of biosensors were optimized, and the detection mechanism was studied. We also detected AChE in serum samples based on TGA-QDs or MGA-QDs. The linear range was 10-140 and 50-1000 U/L, respectively. The excellent performance of this novel biosensor demonstrated that this strategy has prodigious potential to be applied in practice detection of AChE.     

Direct electrochemistry and electrocatalysis of horseradish peroxidase in MnO2 nanosheet film

A novel material MnO2 nanosheet has been used as the support matrix for the immobilization of horseradish peroxidase （HRP）. HRP entrapped in MnO2 nanosheet film exhibits facile direct electron transfer with the electron transfer rate constant of 6.86 s^-1. The HRP/MnO2 nanosheet film gives a reversible redox couple with the apparent formal peak potential （E^0＇） of -0.315 V （vs. Ag/AgCl） in pH 6.5 phosphate buffer solution （PBS）. The formal potential E^0＇ of HRP shifts linearly with pH with a slope of -53.75 mV.pH^-1, denoting that an electron transfer accompanies single-proton transportation. The immobilized HRP shows an electrocatslytic activity to the reduction of H2O2. The response time of the biosensor for H2O2 is less than 3 s, and the detection limit is 0.21 μmol · L^-1 based on signal/noise = 3.     

Influence of particle size and temperature on the dielectric properties of CoFe2O4 nanoparticles

The objective of this study was to establish the dielectric properties of CoFe2O4 nanoparticles with particle sizes that varied from 28.6 to 5.8 nm. CoFe2O4 nanoparticles were synthesized using a chemical coprecipitation method. The particle sizes were calculated accord-ing to the Scherrer formula using X-ray diffraction （XRD） peaks, and the particle size distribution curves were constructed by using field-emission scanning electron microscopy （FESEM） images. The dielectric permittivity and loss tangents of the samples were determined in the frequency range of 1 kHz to 1 MHz and in the temperature range of 300 to 10 K. Both the dielectric permittivity and the loss tangent were found to decrease with increasing frequency and decreasing temperature. For the smallest CoFe2O4 nanoparticle size, the dielectric per-mittivity and loss tangent exhibited their highest and lowest values, respectively. This behavior is very useful for materials used in devices that operate in the microwave or radio frequency ranges.     

Microplate chemiluminescence enzyme immunoassay for the quantitative evaluation of carbohydrate antigen 72-4 in human serum

A highly sensitive and specific microplate chemiluminescence enzyme immunoassay （CLEIA） was developed for the quantitative evaluation of carbohydrate antigen 72-4 （CA72-4） in human serum, using luminol-H2O2 catalyzed by horseradish peroxidase （HRP） as the chemiluminescence system. The simple and quick determination was accomplished through a sandwich reaction mode. Several physicochemical parameters of the immunoreaction, including incubation conditions, antibody coating conditions, dilution ratio of anti-CA72-4-HRP conjugate, and chemiluminescence reaction time, were studied and optimized. The proposed method exhibited a linear range of 0-200 U/mL with correlation coefficient and detection limit of 0.9995 and 0.18 U/mL, respectively. The inter-assay and intra-assay coefficients of variation （CV） were both less than 10%. The average recovery of two clinical sera with low and high concentration CA72-4 was 99.3% and 98.7%, respectively. Normal tumor markers, including AFP, CEA, CA24-2, CA19-9 and CA15-3, did not cross-react with each other. The method＇s stability was evaluated by assessing its analytical performance after storing the immunoreagents at 4℃ and 37℃ for 7 days. Little difference was found, indicating satisfactory stability of the method. The present method has been successfully applied to the detection of CA72-4 human serum, and showed a good correlation with the commercially available ELISA kit （r^2=0.9383）. This method showed great potential in the fabrication of diagnostic kit for CA72-4, and could be well used in diagnosis of cancer in clinical practice.     

Immobilization of Hemoglobin on Platinum Nanoparticles-Modified Glassy Carbon Electrode for H_2O_2 Sensing

This work described an amperometric hydrogen peroxide (H2O2) biosensor based on immobilization of hemoglobin (Hb) on a glassy carbon (GC) electrode modified by platinum nanoparticles, which was prepared by an in situ chemical reductive growth method. The electrochemical impedance measurements confirmed that the Hb was immobilized on the platinum nanoparticles-modified glassy carbon surface and has a synergistic effect with platinum nanoparti-cles in improving the catalytic reduction of H2O2. The Hb immobilized platinum nanoparticles-modified GC (Hb/Pt/GC) electrode displays an effective catalytic response to the reduction of H2O2. A linear dependence of the catalytic current versus H2O2 concentration was obtained in the range of 5.0×10-6 to 4.5×10-4 mol·L-1 with a detection limit (S/N=3) of 7.4×10-7 mol·L-1.     

Study on peptide-peptide interaction using high-performance affinity chromatography and quartz crystal microbalance biosensor

The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography （HPAC） and quartz crystal microbalance （QCM） biosensor. Fragment 1-14 of human interferon-β （hlFN-βwas chosen as sense peptide and its three antisense peptides （AS-IFN 1, AS-IFN 2, and AS-IFN 3） were designed according to the degeneracy of genetic codes. The affinity column was prepared with sense peptide as ligand and the affinity chromatographic behavior was evaluated. Glu-substituted antisense peptide （AS-IFN 3） showed the strongest binding to immobilized sense peptide at pH 7.5. A quartz crystal microbalance-flow injection analysis （QCM-FIA） system was introduced to investigate the recognition process in real-time. The equilibrium dissociation constants between sense peptide and AS-IFN 1, AS-IFN 2 and AS-IFN 3 measured 2.08×10^-4, 1.31×10^-4 and 2.22×10^-5 mol/L, respectively. The mechanism study indicated that the specific recognition between sense peptide and AS-IFN 3 was due to sequence-dependent and multi-modal affinity interaction.     

In-situ secondary growth of nanocube-based Prussian-blue film as an ultrasensitive biosensor

A regular nanostructure has been widely confirmed to result ina marked improvement in material performance in biosensing applications.In the present study,a regular nanostructured Prussian blue(PB) film with two heterogeneous crystal layers was synthesized in-situ using a secondary growth method.A PB seed layer was first controlled to form uniform cube-like crystal nuclei through an ultrasonic reaction with a single reactant.Then,well-defined 100 nm PB nanocubes were further crystallized on this seed layer using a self-assembly approach.In order to accelerate the electron transfer rate during the enzyme reaction for glucose detection,the graphene was used as the main cross-linker to immobilize glucose oxidase on the PB film.The as-prepared biosensor exhibited high electrocatalysis and electron conductivity for the detection of trace glucose with a sensitivity of141.5 μA mM~(-1) cm~(-2),as well as excellent anti-interference ability in the presence of ascorbic acid and uric acid under a low operation potential of-0.05 V.     

Research of the terahertz dual-band notch filter based on FSS structures

In this paper, a dual-band notch filter for two-layer frequency selective surface （FSS） at terahertz （THz） frequency with high fil- tering performance has been realized on high-resistivity silicon substrates. With periodic metallic resonators patterned on the sili- con wafer, the designed filter can provide two tunable resonant frequencies for terahertz application. The transmission response was improved by introducing an extra surrounding pattern around the T-shaped structure, and the location of transmission drop was tuned independently with geometric parameters. Simulated by finite-integral time-domain method, the filter is designed to operate between 0.2 and 0.6 THz with dual-band band-stop performance, a salient feature of this design is making the low sensi- tivity of its frequency response to the incident angles, which allows to place the filter close to the radiation source with spherical wave fronts. The proposed structures were fabricated using photolithography and tested by THz time-domain spectroscopy system Experimental results show that the transmission response has more than 12 and 32 dB rejections near 285 and 460 GHz respectively, which is in good agreement with the simulation result.     

Amplified optical transduction of proteins derived from Mo6S9-xIx nanowires

We demonstrate that Mo6S9-xIx nanowires(MoSI NWs) enable the detection of proteins with cytochrome c as a model protein using UV-vis spectrometry.The association of cytochrome c with the nanowires was verified by scanning electroctron microscopy,X-ray photoelectron,light scattering and micro-FTIR spectroscopies.Our results show that MoSI NWs is a promising nanostructure material for the development of ultrasensitive sensors for detecting proteins.The new MoSI NW derived amplification bioassay is expected to provide a straightforward and effective strategy for protein analysis and biosensor construction.     

Rapid detection of Cyfra 21-1 by optical-biosensing based on chemiluminescence immunoassay using bio-functionlized magnetic nanocomposites

This letter reports a chemiluminescene immunoassay method combined with immunomagnetic separation to rapidly detect Cyfra 21-1, in which bio-functionlized magnetic nanocomposites were used as mobile substrate for capturing and isolating the cyfra 21-1 proteins. After the captured Cyfra 21-1 further reacted with horseradish peroxidase-conjugated anti-Cyfra 21-1 antibody to form a sandwich immunocomplex, the chemiluminescence would be produced as a result of addition of the chemiluminescent substrate. A home-made optical biosensor was designed to detect the chemiluminescence instead of other large instruments. There is a good linear response between the chemiluminescence intensity and the concentration of Cyfra 21-1 in the range from 0.2 to 50 ng/mL. The whole detection process including incubation, washing and detection could be performed within 45 min. The proposed method offers a simple, noninvasive and reliable tool for detecting non-small cell lung cancer and has potential application for clinical testing.     

Rod-shaped Au@PtCu nanostructures with enhanced peroxidase-like activity and their ELISA application

Pt and its based alloy nanoparticles （NPs） have been reported to demonstrate novel enzyme-like activities. Varying composition is very important to realize the opti- mization of their functions through the tuning of electronic structure. In this paper, our effort is focused in this direction by tailoring the electronic structure of Pt NPs via alloying with copper. Using gold nanorod （Au NR） as core, a simple method to prepare PtCu alloy shell is developed （termed as Au@PtCu NR）. The introduction of copper could result in endcap-preferred growth mode owing to the lattice mismatch between alloy shell and the Au core. The variation in the electronic structure changes the substrate affinity, and enhanced affinity was found for H202. Besides, the designed Au@PtCu nanostructures have realized spatial separation of catalytic and recognition sites. Binding of recognition antibodies had negligible effect on their catalytic activity. Based on their peroxidase- like activity, a highly sensitive detection of human immunoglobulin G （IgG） was demonstrated in a direct enzyme-linked immunosorbent assay （ELISA） mode. The detection limit can be as low as 90 pg/mL.     

Antibacterial Cellulose Coated with N-halamine by Pad-Cure Process

In order to improve the antibacterial property of cellulose,a new N-halamine antibacterial material precursor was synthesized.1,2,3,4-Butanetetracarboxylic acid( BTCA) was used to attach the N-halamine precursor onto cotton fabric as the cross-linking agent. The synthesized compound was characterized by1 H nuclear magnetic resonance(1H NMR). The cotton fabric treated with Nhalamine precursor was characterized by Fourier transform infrared spectroscopy( FT-IR) and scanning electron microscopy( SEM).The antimicrobial efficacy, washing durability and UVA light stability were investigated. The cotton fabric treated with the Nhalamine precursor could be rendered biocidal after exposure to dilute household bleach. The chlorinated cotton fabric shows great antimicrobial efficacy,100% of Staphylococcus aureus( S. aureus)with 1.00 ×10~7 CFU and 99. 998% of Escherichia coli (E. coli)O157∶H7 with 2. 20 × 10~7 CFU can be inactivated with 30 min of contact. Washing durability tests indicate that over 55% of the chlorine can be regained upon rechlorination after 50 washing cycles,and UVA light stability tests show that over 62% of the chlorine can be regenerated after irradiation of 12 h.     

Motion Planning Method for Obstacle Avoidance of 6-DOF Manipulator Based on Improved A* Algorithm

The conventional A* algorithm may suffer from the infinite loop and a large number of search data in the process of motion planning for manipulator. To solve the problem,an improved A* algorithm is proposed in this paper by the means of selecting middle points and applying variable step segments searching during the searching process. In addition,a new method is proposed for collision detection in the workspace. In this paper,the MOTOMAN MH6 manipulator with 6-DOF is applied for motion plan. The algorithm is based on the basis of the simplification for the manipulator and obstacles by cylinder enveloping. Based on the analysis of collision detection,the free space can be achieved which makes it possible for the entire body to avoid collisions with obstacles. Compared with the Conventional A*,the improved algorithm deals with less searching points and performs more efficiently. The simulation developed in VC + + with OpenGL and the actual system experiments prove effectiveness and feasibility of this improved method.     

Optimal design of label-free silicon“ lab on a chip” biosensors

This paper reported the optimal design of label-free silicon on insulator(SOI) "lab on a chip"biosensors.These devices are designed on the basis of the evanescent field detection principles and interferometer technologies.The well-established silicon device process technology can be applied to fabricate and test these biosensor devices.In addition,these devices can be monolithically integrated with CMOS electronics and microfuidics.For these biosensor devices,multi-mode interferometer(MMI) was employed to combine many stand-alone biosensors to form chip-level biosensor arrays,which enable realtime and label-free monitoring and parallel detection of various analytes in multiple test samples.This sensing and detection technology features the highest detection sensitivity,which can detect analytes at extremely low concentrations instantaneously.This research can lead to innovative commercial development of the new generation of high sensitivity biosensors for a wide range of applications in many fields,such as environmental monitoring,food security control,medical and biological applications.