双通道SH—APM生物传感器对尿浓度检测的研究

以双通道声板波器件（ｓｈａｒ ｈｏｒｉｚｏｎｔａｌ ａｃｏｕｓｔｉｃ ｐｌａｔｅｍｏｄｅ，ＳＨ－ＡＰＭ）作为换能器对生物量进行测量的方法，解决了生物传感 器中普遍存在由于电极与生物易感物质直接接触而造成的不良后果，同时克服了常用单通道声板波传感器进行生物量测试的缺点。系统将该器件与生物易感物质（尿酶）结合组成的生物传感系统对尿的浓度进行测试。声板波器件的中心频率为４９．６６ＭＨz。

The biofilm characteristics and enhanced performance of a marine microbial-electrolysis-cell-based biosensor under positive anodic potential

Microbial fuel cells have already been used as biosensors to monitor assimilable organic carbon(AOC).However,their signal production from AOC is known to be completely suppressed by dissoved oxygen(DO).In this study,two identical microbial electrolysis cell(MEC)based biosensors were inoculated with marine sediment and operated at two different anodic potentials,namely-300 mV and+250 mV relative to Ag/AgCl.The MEC biosensor operated under positive anodic potential conditions had electrochemically active microbial communities on the anode,including members of the Shewanellaceae,Pseudoalteromonadaceae,and Clostridiaceae families.However,the strictly anaerobic members of the Desulfuromonadaceae,Desulfobulbaceae and Desulfobacteraceae families were found only in the negative anodic potential MEC biosensor.The positive anodic potential MEC biosensor showed several other advantages as well,such as faster start-up,significantly higher maximum current production,fivefold improvement in the AOC detection limit,and tolerance of low dissolved oxygen,compared to those obtained from the negative anodic potential MEC biosensor.The developed positive anodic potential MEC biosensor can thus be used as a real-time and inexpensive detector of AOC concentrations in high saline and low DO seawater.

Mushroom Pulp Tissue-Based Membrane-Ferrocene-Modified L-Tyrosine Biosensor

A new approach for assembling amperometric mushroom pulp tissue based membrane electrode for determination of L tyrosine analysis is proposed. Ferrocene is used as a mediator of electron transfer between tyrosinase in mushroom tissue and a graphite electrode. The optimal operation conditions are studied. The linear response range of the biosensor is 2 0×10 -4 to 4 5×10 -3 mol·L -1 with response time of less than 5 min and lifetime of at least 30 d. The biosensor can be applied to practical sample analysis.

Determination of mercury and nickel by amperometric biosensor prepared with thermostable lactate dehydrogenase

Response of biosensor prepared with the thermostable bacterial LDH enzyme was analyzed in the presence of mercury and nickel.For electrode preparation,the enzyme was purified and immobilized on a gold sheet coated by PGA-pyrrole polymeric material.The working electrode was tested at increasing concentration of lactate in the presence of two different concentrations of mercury and nickel.Current response of biosensor decreased from 0.32 μA to 0.09 μA and 4.13 μA to 2.63 μA when 25×10-7 mmol/L mercury and 17×10-5 mmol/L nickel were included in the working solution,respectively.Sensitivity of the electrode decreased from 0.010 2 μA/（mmol·L-1） to 0.0043 μA/（mmol·L-1） in the presence of 25×10-7 mmol/L mercury.On the other hand,the presence of nickel did not result in a decrease in electrode sensitivity.The results pointed out that the prepared biosensor is useful to detect mercury in a sample containing both mercury and nickel together.

New amperometric glucose biosensor by entrapping glucose oxidase into chitosan/nanoporous ZrO_2/multiwalled carbon nanotubes nanocomposite film

A new nanocomposite material for construction of glucose biosensor was prepared. The biosensor was formed by entrapping glucose oxidase(Gox) into chitosan/nanoporous ZrO2/multiwalled carbon nanotubes nanocomposite film. In this biosensing thin film, the multiwalled carbon nanotubes can effectively catalyze hydrogen peroxide and nanoporous ZrO2 can enhance the stability of the immobilized enzyme. The resulting biosensor provides a very effective matrix for the immobilization of glucose oxidase and exhibits a wide linear response range from 8 μmol/L to 3 mmol/L with a correlation coefficient of 0.994 for the detection of glucose. And the response time and detection limit of the biosensor are determined to be 6 s and 3.5 μmol/L, respectively. Another attractive characteristic is that the biosensor is inexpensive, stable and reliable.

Study on Piezoelectric Immunosensor for the Detection of H9-subtype Avian Influenza Virus

[Objective] The aim is to develop the piezoelectric immunosensor to detect H9-subtype avian influenza virus（AIV）.[Method] The immunosensor chip was constructed by self-assembling mercaptopmpionic acid（MPA） to be monolayer on the silver-coated electrode of quartz crystal and coupling the monoclonal antibody to H9 subtype AIV with N-ethy-N＇-（3-dimethyl aminopropyl）carbodiimide hydrochloride（EDC） and N-hydroxysuccinimide（NHS）.The immunosensor to detect H9 subtype AIV was established.[Result] The results showed that the immunosensor displayed better specificity to H9 AIV and had no response to H5AIV and NDV when it was used for detection.The sensitivity test indicated the detection sensitivity for the H9 subtype AIV could reach 20-100 EID50.[Conclusion] The research provided a foundation for further research on the immunosensor for detecting AIV and it could be a new approach to detect other related viruses.

Amino Acid Sensing Using an Ion-Sensitive Field-Effect Transistor

Ion-sensitive field-effect transistor （ISFET）-based biosensor for amino acids was proposed, and the response of the sensor to amino acids was evaluated. As a molecular recognition element, aminoacyl-tRNA synthetase which would be expected to have high selectivity for the corresponding amino acids was used, aminoacyl-tRNA synthetase was coated onto an ISFET electrode, and the response of the biosensor to amino acids was evaluated. The amino acid sensor for tyrosine showed a selective response to tyrosine because of the specific binding ability of aminoacyl-tRNA synthetase for tyrosine and from 300μM to 900 μM of tyrosine could be measured （r2 〉 0.969）.

Novel biosensor-based microarray assay for detecting rs8099917 and rs12979860 genotypes

AIM:To evaluate a novel biosensor-based microarray(BBM) assay for detecting rs12979860 and rs8099917 genotypes.METHODS:Four probes specific for rs8099917C/T or rs12979860G/T detection and three sets of quality control probes were designed,constructed and arrayed on an optical biosensor to develop a microarray assay.Two sets of primers were used in a one tube polymerase chain reaction(PCR) system to amplify two target fragments simultaneously.The biosensor microarray contained probes that had been sequenced to confirm that they included the rs8099917C/T or rs12979860G/T alleles of interest and could serve as the specific assay standards.In addition to rehybridization of four probes of known sequence,a total of 40 clinical samples collected from hepatitis C seropositive patients were also tested.The target fragments of all 40 samples were amplified in a 50 μL PCR system.Ten μL of each amplicon was tested by BBM assay,and another 40 μL was used for sequencing.The agreement of the results obtained by the two methods was tested statistically using the kappa coefficient.The sensitivity of the BBM assay was evaluated using serial dilutions of ten clinical blood samples containing 10 3-10 4 white cells/μL.RESULTS:As shown by polyacrylamide gel electrophoresis,two target segments of the interleukin 28Bassociated polymorphisms(SNPs) were successfully amplified in the one-tube PCR system.The lengths of the two amplified fragments were consistent with the known length of the target sequences,137 and 159 bps.After hybridization of the PCR amplicons with the probes located on the BBM array,the signals of each allele of both the rs8099917 SNPs and rs12979860 SNPs were observed simultaneously and were clearly visible by the unaided eye.The signals were distinct from each other,could be interpreted visually,and accurately recorded using an ordinary digital camera.To evaluate the specificity of the assay,both the plasmids and clinical samples were applied to the microarray.First,30 PCR amplicons of the various SNP alleles were hybridized on the BBM microarray.Full agreement between plasmids and the BBM assay was observed,with 30/30 correct matches(100%).The kappa value for the BBM assay with plasmids was 1.00(P < 0.05).For the 40 clinical blood samples,the BBM assay hybridization and direct sequencing results were compared for each amplicon.For patient blood samples,agreement was 28/28 for rs8099917T/T,9/11 for rs8099917T/G,1/1 for rs8099917G/G,24/24 for rs12979860C/C,11/14 for rs12979860C/T,and 2/2 for rs12979860T/T.Only five clinical samples of amplicon assay and direct sequencing results were discordant and heterozygotes:2/11 rs8099917T/G and 3/14 rs12979860C/T.The agreement of outcomes between BBM assay and direct sequencing for the detection of rs8099917 and rs12979860 was 95% and 92.5%,respectively;and the corresponding kappa values were 0.88 and 0.85(A kappa value > 0.75 was defined as substantial agreement).The BBM assay and sequencing had similar specificities for detection and identification of the two SNPs and their alleles.The sensitivity evaluation showed that the BBM assay could detect and identify SNP sequences present in blood samples containing as few as 10 2 white blood cells/μL.CONCLUSION:This biosensor microarray assay was highly specific,sensitive,rapid and easy to perform.It is compatible with clinical practice for detection of rs8099917 and rs12979860.

Polyaniline-graphite composite film glucose oxidase electrode

A novel polyaniline-graphite composite film glucose oxidase （PGCF GOD） electrode was developed. The PGCF was synthesized by cyclic voitammetry method in 0.5 mol/L H2SO4 solution containing 1 g/L graphite powder and 0.2 mol/L aniline. The PGCF GOD electrode was prepared by doping GOD into the composite film. The morphology of the PGCF and the response property of the PGCF GOD electrode were investigated by scanning electron microscopy and electrochemical measurement, respectively. The results show that the PGCF has a porous and netty structure and the PGCF GOD electrode has excellent response property such as high sensitivity and short response time. Influences of pH value, temperature, glucose concentration and potential on the response current of the electrode were also discussed. The sensor has a maximum steady-state current density of 357.17μA/cm2 and an apparent Michaelis-Menten constant of 16.57 mmol/L. The maximum current response of the enzyme electrode occurs under the condition ofpH 5.5, 0.8 V and 65℃.

AUTHENTICATED SYMMETRIC-KEY ESTABLISHMENT FOR MEDICAL BODY SENSOR NETWORKS

This study concerns security issues of the emerging Wireless Body Sensor Network (WBSN) formed by biomedical sensors worn on or implanted in the human body for mobile healthcare appli-cations. A novel authenticated symmetric-key establishment scheme is proposed for WBSN,which fully exploits the physiological features obtained by network entities via the body channel available in WBSN but not other wireless networks. The self-defined Intrinsic Shared Secret (ISS) is used to replace the pre-deployment of secrets among network entities,which thus eliminates centralized services or au-thorities essential in existing protocols,and resolves the key transport problem in the pure symmet-ric-key cryptosystem for WBSN as well. The security properties of the proposed scheme are demon-strated in terms of its attack complexity and the types of attacks it can resist. Besides,the scheme can be implemented under a light-weight way in WBSN systems. Due to the importance of the ISS concept,the analysis on using false acceptance/false rejection method to evaluate the performance of ISS for its usage in the scheme is also demonstrated.

Laccase biosensor using magnetic multiwalled carbon nanotubes and chitosan/silica hybrid membrane modified magnetic carbon paste electrode

A simple and rapid strategy to construct laccase biosensor for determination of catechol was investigated. Magnetic multiwalled carbon nanotubes (MMCNT) which possess excellent capability of electron transfer were prepared by chemical coprecipitation method. Scanning electron microscope (SEM) and vibrating sample magnetometer (VSM) were used to identify its surfacetopography and magnetization, respectively. Laccase was immobilized on the MMCNT modified magnetic carbon paste electrode by the aid of chitosan/silica (CS) hybrid membrane. Using current-time detection method, the biosensor shows a linear response related to the concentration of catechol in the range from 10-7 to 0.165×10-3 mol/L. The corresponding detection limit is 3.34×10-8 mol/L based on signal-to-noise ratios (S/N) ≥3 under the optimized conditions. In addition, its response current retains 90% of the original after being stored for 45 d. The results indicate that this proposed strategy can be expected to develop other enzyme-based biosensors.

Determination of the Antioxidant Capacity of Different Types of Bread and Flour Using a Superoxide Dismutase Biosensor

The aim was to experimentally evaluate the antioxidant capacity of different types of bread and of the relative flour used for bread production utilizing a superoxide dismutase （SOD） biosensor recently developed by the present authors. Measurements were carried out by comparing the biosensor response to the concentration of superoxide radical produced in solution using a xanthine/xanthine oxidase system in the presence and in the absence of the antioxidant sample considered, respectively. Precision of antioxidant capacity measures for crust and crumb of the different breads was found to be good （RS D% ≤ 8%） and acceptable for the watery suspension and filtrate of the different flours studied （RSD% ≤ 12%）. The obtained results indicated that general flours show higher antioxidant capacity values than the corresponding breads and that crusts show always an antioxidant capacity definitely larger than the crumb. Lastly, the antioxidant capacity values were compared with those of almond, red pepper and strawberry, three foods containing powerful natural antioxidants.

Self-Assembled Monolayer of Lipoic Acid on Gold and Its Application to Rapid Determination of 2, 3, 7, 8-Tetrachlorodibenzo-p-Dioxin

Lipoic acid (LA) has received great attention in the area of gold surface functionalization. In this study, LA was employed as a linker for the attachment of antibody to the gold surface of surface plasmon resonance (SPR) chip. By using this chip in a homemade SPR immunosensor, low molecular weight compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) can be detected at a low level of 0.01 ng/mL. There is a good linear relationship(R2 =0.943 1) between the results of SPR biosensor and enzyme-linked immunosorbent assay(ELISA).

A Graphene Oxide-based Immuno-biosensor for Vibrio parahaemolyticus Detection

Vibrio parahaemolyticus is the leading causal agent of human acute gas- troenteritis. Real-time accurate detection means is the key to prevention and control of its spread. This study provided a novel detection strategy for realizing rapid and specific determination of V. parahaemolyticus by labeling its monoclonal antibody （Ab） with quantum dots （QDs）. The results showed that the fluorescence of these QDs-Ab bioconjugates was quenched by graphene oxide （GO） to produce a bacteri- um capture probe. And the optimal quenched concentration of GO was 60 ng/ml. When the bacterium capture probe was exposed to the target, green color fluores- cence was turned on by releasing the QDs-Ab due to the antibody antigen combi- nation. The detection limit of V. parahaemolyticus was 104 CFU/ml based on 3 times signal-to-noise ratio. The specificity of the FRET sensor towards V. para- haemolyticus was examined by comparing with controls such as V. splendidus, V. alginolyticus, Edwardsiella tarda and Aeromonas hydrophila with the same condition. The controls couldn＇t cause obvious fluorescence alteration, while the target resulted in significant fluorescence enhancement. This strategy could be further used as a universal method for any bacterial determination by changing the conjugated antibod- ies in early disease diagnosis. Therefore, the sensor has good potential to expand its application to the early diagnosis and determination of bacteria.

Fiber optic biosensor of immobilized firefly luciferase

Luciferase from firefly lantern extract was immobilized on CNBr activated Sepharose 4B. The kinetic properties of immobilized luciferase were extensively studied. The K m′ for D luciferin is 11.9 μmol/L, the optimum pH and temperature for Sepharose bound enzyme were 7.8 and 25℃ respectively. A luminescence fiber optic biosensor, making use of immobilized crude luciferase, was developed for assay of ATP. The peak light intensity was linear with respect to ATP concentration in range of 10 -9 -10 -5 mol/L. A biological application was also demonstrated with the determination of serum ATP from rats bred in low versus normal oxygen environments.

Low Noise Readout Circuit for Biosensor SoC

Presented is a low noise interface circuit that is tuned to the needs of self-assembly monolayers biosensor SoC. The correlated double sampling(CDS) unit of the readout circuit can reduce 1/f noise, KTC noise and fixed noise of micro arrays effectively. The circuit is simulated in a 0.6 μm/level 7 standard CMOS process, and the simulated results show the output voltage has a good linearity with the transducing current of the micro arrays. This is a novel circuit including four amplifiers sharing a common half-circuit and the noise reducing CDS unit. It could be widely used for micro array biosensors.

A ferrocene-mediated anti-interfering glucose biosensor based on glutin and cellulose acetate

A ferrocene-mediated glucose biosensor removing interference of ascorbic acid and uric acid was developed by coating of ferrocene, glutin and cellulose acetate on screen-printed gold electrode surface. The resuhs show that it can detect glucose sensitively in the presence of uric acid and ascorbic acid, and also suppress the leakage velocity of fcrrocene. Compared to the currents of the pretreated electrode, it decreases the current of uric acid and ascorbic acid by 99.4% and 98.8% at 400 mV, respectively, with a dynamic range of 0 - 30 mM for glucose, sensitivity of 30.73 nA/mM, response time of 10 s, and correlation coefficient of 0.998 8.

Remote sensing Extraction model of redtide biomass by airborne hyperspectral technique

Our work is based on the known research results of inherent optical quality of ocean color constituents.According to optimized parameters and induced fluorescence term of chlorophyll, this paper puts forward a remote sensing reflectance model of sea water, which is fitted in Liaodong Bay of Bohai. An inverse model that can evaluate redtide biomass according to chlorophyll retrieval is provided by inducing a functional extreme problem. The calculation example of the model indicates that the inversion model has explicit mathematic and physical meaning, but its practicability needs to be verified.

Toxicity of single-walled carbon nanotubes on green microalga Chromochloris zofingiensis

Nanoparticles,or particles in size of 1-100 nm,are extensively used in the world in different applications.For instance,single-walled carbon nanotubes(SWCNTs) are commonly used in consumer products,such as biosensors,drug and vaccine delivery transporters,and novel biomaterials.Although nanoparticles do not cause safety concerns to consumers who use nanoparticle-containing products,these small particles are potentially harmful for workers who produce them in factories or in cases of discharge to aquatic ecosystems.SWCNTs do not have a natural analogue,so the effects on health of their disposal remain largely unknown.In this study,we evaluated the effects of SWCNTs on a population of the green microalga Chromochloris zofingiensis and the profile and production of pigments and fatty acids.The alga was incubated with SWCNTs for 6 days in 0(control),40,80,160,or 320 mg/L concentrations.SWCNTs showed both positive and negative effects on the growth of C.zofingiensis,with a biomass enhancement at low levels(40-160 mg/L) but inhibition at high levels(320 mg/L).By contrast,a decreased accumulation of fatty acids and pigments of C.zofingiensis was observed over the range of the tested concentrations.These results indicate that the markers on the inhibitive toxicity of SWCNTs are increasingly sensitive in the following order:biomass and fatty acids < primary carotenoids < chlorophylls < secondary carotenoids.C.zofingiensis is a suitable microalga for evaluating the ecotoxicological hazards of SWCNTs,especially in terms of pigmentation response.

Bacteriorhodopsin and SWCNT Scaffold for Optical Nanobiosensor

This paper describes theoretical steps to develop an optical nanobiosensor using bacteriorhodopsin （BR） as the biomembrane and Single-Walled Carbon NanoTube （SWCNT） as the scaffold. Bacteriorhodopsin is a retinal protein used by archaea that come under the family of halobacteria. This retinal protein acts as a proton pump and resulting proton gradient is used to change the voltage that pass across the drain and source. The biosensor contains nano ISFET where the channel is made of a carbon nanotube for the conduction of current. The gate is replaced by bacteriorhodopsin biomembrane. Bacteriorhodopsin can be used as a molecular-level ultra fast bi-stable red / green photo switch for making 3D optical molecular memories that reliably store data with 10,000 molecules/bit. The molecules switch in femtoseconds. Biomembrane will sense 510 nm and 650 nm wavelength of light and the sensing voltage can be used to convert the data into digital signals. This molecular level memory device can be used for ‘Read-Write＇ operations. The sensor performance will also be ultra fast since it uses photons for the data storage, which are much faster than electrons used in normal memory devices, and the 3D storage capacity is much higher maximum of 10^13/cm^2.