基于纳米磁珠修饰印刷电极的牛奶中三聚氰胺检测安培免疫传感器

共固定氯代邻菲哕啉双核铜配合物(Cu2(phen)2Cl2(μ-Cl)2,简称CuzL)和包被三聚氰胺抗体(anti MA)的FbO4/Au胶(简称纳米磁珠)于丝网印刷电极(SPCE)表面,构建了一种测定牛奶中三聚氰胺(简称MA)含量的安培免疫传感器.当该传感器在含MA溶液中温育时,MA和同时加入的HRP标MA二抗(HRP-anti MA Ⅱ),与电极表面的anti MA形成三元免疫复合物.该复合物上的HRP能催化HzO2还原,并经过Cu2L传导在电极表面形成电流,电流强Ⅰ与MA浓度在0.5～40和60～100ng/mL范围内呈线性关系,检测限为0.25 ng/mL.在pH 6.2的PBS底液中,该传感器制作简单、可抛弃,对MA检测时间小于20 min,灵敏度为1μA·ng-1mL,明显高于酶联免疫吸附(ELISA)法;可实现牛奶中MA免分离现场分析.     

基于双正交多小波的红外和可见光图像融合

利用双正交单小波的对称性和良好的滤波性能，给出了具有GMPs (Good Multifilter Properties)的双正交多小波滤波器，对其经过平衡处理后，结合Burt提出的选择和平均的融合方案，对可见光图像和红外图像进行了融合实验，充分利用红外波段和可见光波段的信息互补性，达到便于观察的目的。仿真结果表明该方法能取得较好的融合效果。     

血清C-反应蛋白在小儿支原体肺炎鉴别诊断中的临床观察

目的:观察血清C-反应蛋白(CRP)水平在小儿细菌性肺炎和支原体肺炎中的临床改变,探讨提高小儿支原体肺炎诊断符合率的检查方法方法:选择确诊为肺炎的患儿158例,根据其他实验室检查及临床疗效,分为细菌感染组82例和支原体感染组76例,比较两组患者就诊时血清CRP水平及治疗过程中血清CRP改变情况结果:细菌感染组与支原体感染组血清CRP阳性例数和就诊时血清CRP水平比较,差异具有高度统计学意义(P＜0.01);细菌感染组惠儿血清CRP水平与治疗天数呈正相关性(r=0.833,P＜0.01)结论:在小儿肺炎的诊断及治疗过程中,应积极行血清CRP,可提示患儿肺炎类型并指导临床治疗.     

电化学极化玻碳电极构建的葡萄糖生物传感器

在硝酸溶液中电化学极化处理玻碳电极(GCE),以硫堇(TH)为电子介体,通过金-硫、金-氮共价键作用和静电吸附作用将纳米金(GNPs)、葡萄糖氧化酶(GOD)和辣根过氧化物酶(HRP)修饰于电极上,最后在电极上滴涂Nafion水溶液制备抗干扰膜并固定酶电极,构建了一种新型双酶葡萄糖生物传感器.实验结果表明:电化学极化处...     

基于体系结构依赖图的Wright形式化语言的切片方法

将切片技术应用于软件体系结构中,根据程序依赖图和系统依赖图的构建思想,提出了体系结构元素依赖图(SADG)的概念,并给出了构建方法以及基于依赖图的切片算法。利用此切片技术从已有的软件体系结构的Wright描述中,提取使用者感兴趣的软件元素(例如,构件和连接体),从而从体系结构的角度,更好的支持了软件理解、软件维护、软件再工程以及软件复用。     

基于碳纳米管固定葡萄糖氧化酶的ECL葡萄糖传感器的制备

将葡萄糖氧化酶(GOD)固定在多壁碳纳米管(MWCNTs)修饰电极(ME)上,GOD催化氧化葡萄糖生成过氧化氢,并使鲁米诺产生电致化学发光(ECL),据此构建了一种新型ECL葡萄糖传感器.结果表明:CNTs修饰的电极对鲁米诺和H2O2反应具有显著的电催化活性和增敏效果.该传感器对葡萄糖检测的线性范围为0.01～10.0...     

业务层框架构建解决方案的研究

业务逻辑的千变万化,使信息系统的开发和维护都变得越来越困难,如何构建灵活高度可扩展性的业务层便成为亟待解决的问题。本文主要介绍一种灵活、高度可扩展的业务框架的构建方法:提出按照服务(Service)的概念划分业务功能,按照Domain Model—Service—Control思想实现服务导航(Service Direction),采用面向方面的编程方法(AOP)将服务导航从服务的实现中分离,而所有这些均在业务容器(Business Container)里执行,从而实现了业务层相对于表示层的透明化。     

动态角色体系结构与算法研究

引入新一代动态角色系统概念构建VR仿真场景.首先提出了动态角色层概念,解决了传统方法中存在的接口混乱,资源浪费等问题;然后,通过虚拟环境编辑器管理和构建虚拟角色,将其摆放到合适的位置.并具体介绍了动态角色体系结构(动态角色库管理器、动态角色代理器以及工程项目)和工作流程,系统中属性库更新通过OSG(Open Scene Graph)编程实现(给出了实例飞机飞行计划模拟的方法和流程图),软件研发成果可以通过这样的动态角色系统得到很好的保障.     

脂肪醛酮的沸点及色谱保留指数定量构效关系的研究

应用密度泛函理论(DFT),在6-31+G(d)水平上对脂肪醛酮的分子结构几何优化和量化计算.将量子化学计算的部分结果引入分子的拓扑结构中,构建新型分子量子拓扑指数(NQT),然后研究脂肪醛酮的沸点量子拓扑化学的变化规律,同时将Nqr与Gibbs自由能(G),恒容摩尔热容(Cv),配分函数(1gQ)结合起来建立脂肪醛酮色谱保留指数的定量模型.其沸点模型相关系数R>0.99,色谱保留指数模型相关系数R>0.98,并用随机抽样法、留一法(LOO)验证,证明所建模型稳健、合理、有效.本文将量子化学方法与拓扑学方法结合起来,使拓扑方法中某些经验数值上升为理论数值,从而推动拓扑化学的进步.该方法有望在物质的QSPR/QSAR研究中推广.     

叶枝分辨的决策树研究

为了解决当前构建决策树算法中准确度与计算量之间的矛盾,根据粒计算理论,提出了利用叶枝粒之比的决策树构建算法:将条件属性按属性值划分为若干基本粒作为基本分配单位(叶重量),而将基本粒所对应的决策属性值的类别数量作为基本接受单位(枝数量),直接利用叶枝粒比判断确定划分属性,由所选划分属性自顶向下构造决策树.通过理论与实例分析结果表明了该算法简洁且准确性高.     

Ionic liquid and nanogold-modified immunosensing interface for electrochemical immunoassay of hepatitis B surface antigen in human serum

A mediator-free electrochemical immunoassay protocol based on a disposable immunosensor for the detection of hepatitis B surface antigen (HBsAg) in human serum was developed. To fabricate such an immunosensor, a layer of sol–gel composite film containing room temperature ionic liquid and chitosan was initially formed on a glassy carbon electrode. Nanogold particles were then adsorbed onto the membrane via the amine groups of chitosan molecules, and then horseradish peroxidase (HRP)-labeled hepatitis B surface antibodies (HRP-anti-HBs) were immobilized onto the nanogold surface. With a non-competitive immunoassay format, the antibody–antigen complex could be formed by a simple one-step immunoreaction between the immobilized HRP-anti-HBs and HBsAg in sample solution. The formed immunocomplex inhibited partly the active center of the HRP, which decreased the immobilized HRP toward the reduction of H₂O₂. The performance and factors influencing the performance of the immunosensor were evaluated. Under optimal conditions, the current change obtained from the carried HRP relative to H₂O₂ system was proportional to HBsAg concentration in the range of 1.5–400 ng/mL with a detection limit of 0.5 ng/mL (at 3δ). The reproducibility, selectivity, and stability of the proposed immunosensor were acceptable. Moreover, the proposed immunosensors were used to analyze HBsAg in human serum specimens. Analytical results of clinical samples suggested that the developed immunosensor has a promising alternative approach for detecting HBsAg in the clinical diagnosis.     

基于抗体包被纳米金和铜配合物共固定修饰电极的癌胚抗原安培免疫传感器研究

研制了基于抗体包被化学镀纳米金(AuNPs)和[Cu(bpy)2(ONO)]NO3配合物(CuL)共固定修饰玻碳电极(GCE)的安培免疫传感器,并用于血清中癌胚抗原(CEA)的检测。首先将GCE电极表面氧化形成羧基,进而键合上乙二胺。将此胺化电极浸泡在CuL和化学镀金溶液后,CuL可通过π-π堆积作用吸附到GCE表面,并在电极表面还原成30~50nm的纳米金层(GCE|CuL/AuNPs)。将上述电极浸泡在CEA抗体(anti CEA)溶液中,利用AuNPs固定anti CEA,并通过辣根过氧化物酶(HRP)封闭剩余的AuNPs位点,由此构建了一类快速检测CEA的无试剂安培免疫传感器(GCE|CuL-AuNPs/anti CEA-HRP)。其中CuL作为电子媒介体对过氧化脲(CP)有催化还原作用,而且HRP可增强这种作用。当该传感器在37℃下,含CEA的pH6.5PBS溶液中温育30min后,随着温育液中CEA浓度的增加,电极表面形成的免疫复合物也增加,导致CuL对CP的催化电流下降。电流下降百分比1%与CEA浓度在0.1~80ng/mL之间成线性关系,检测限为0.052ng/mL(3σ)。由于采用化学镀法可...     

一种基于纳米金／石墨烯／普鲁士蓝（PB）修饰玻碳电极非标记免疫传感器

采用电聚合的方法将普鲁士蓝（PBl聚合在玻碳电极表面,再将石墨烯修饰在PB上面,然后再采用电沉积的方法将HAuCL直接还原成纳米金粒子,沉积在石墨烯表面,最后将羊抗人IgG抗体直接固定于该修饰的玻碳电极表面,制备了用于人IgG抗原检测的非标记电化学免疫传感器。利用循环伏安法和交流阻抗研究了修饰电极表面的电化学特性,用差分脉冲伏安法对人IgG抗原进行了测定。实验表明,此免疫传感器在含不同浓度人IgG的PBS溶液（pH6．98）中测定,响应电流与人IgG浓度在5．55～455．5ng／mL范围内有良好的线性关系,其相关系数r=0．9926,检测限为0．015ng／mL（S／N=3）。该免疫传感器具有制备简单、响应时间快（5min）、稳定性好等特点。     

Highly sensitive label-free immunosensor for ochratoxin A based on functionalized magnetic nanoparticles and EIS/SPR detection

A label-free immunosensor for the detection of ochratoxin A (OTA) based on use of magnetic nanoparticles (MNPs) was developed. A gold electrode was modified using bovine serum albumin conjugate with a glutaraldehyde-thiolamine linker, creating a layer that prevents non-specific binding of OTA on gold. The OTA antibodies were attached to MNPs using the carbodiimide chemistry and afterwards were immobilized on the modified gold electrode using a strong magnetic field. Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and surface plasmon resonance (SPR) were used to characterize each step in immunosensor development. The impedance variation due to the specific antibody-OTA interaction was correlated with the OTA concentration in the samples. The increase in electron-transfer resistance values was proportional to the concentration of OTA on a linear range between 0.01 and 5 ng/mL, with a detection limit of 0.01 ng/mL. SPR measurements showed a larger response range (1-50 ng/mL) with a detection limit of 0.94 ng/mL. Analytical results were in accordance with standard ELISA test kit.     

基于囊状银钯铂合金的无标记型癌胚抗原免疫传感器的研制

该文将所合成的中空囊状银钯铂合金和壳聚糖（CHIT）修饰于玻碳电极（GCE）表面,利用中空囊状银钯铂合金比表面积大、反应活性位点多、导电能力良好、催化性能优异和吸附能力强等优点将癌胚抗原的抗体（anti-CEA）固定到电极表面,从而制得高灵敏的无标记型癌胚抗原（CEA）免疫传感器。当抗体与抗原发生免疫反应形成复合物时,会降低银钯铂合金的电催化活性并增加传质阻力。采用示差脉冲伏安法（DPV）检测电化学探针铁氰化钾的响应电流信号的减小程度,实现对CEA的检测。实验考察了电极表面的电化学行为,并对免疫传感器的性能进行了研究。在最优的实验条件下检测癌胚抗原的线性范围为0.5-80.0 ng/mL,线性相关系数为0.989 0,检测下限为0.17 ng/mL。     

基于Nafion-碳纳米管-球形二茂铁甲酸纳米复合物构建的电流型人绒毛膜促性腺激素免疫传感器的研究

该文采用超声溶剂置换方法合成球形二茂铁甲酸纳米粒子,以Nafion分散的多壁碳纳米管与球形二茂铁甲酸形成纳米复合物,并将该复合物固定在玻碳电极（GCE）表面,用EDC/NHS将二茂铁甲酸与L-半胱氨酸（L-Cys）交联,形成带有巯基官能团的复合膜,再键合沉积的纳米金（nano-Au）,并通过纳米金吸附人绒毛膜促性腺激素抗体（anti-HCG）,最后用牛血清蛋白（BSA）封闭电极上的非特异性吸附位点,从而制得人绒毛膜促性腺激素（HCG）免疫传感器。实验结果表明,该免疫传感器的响应电流与HCG的浓度有良好的线性关系,其线性范围为0.05 mIU/mL-200 mIU/mL,检出限为0.015 mIU/mL。     

Alpha-1-fetoprotein antibody functionalized Au nanoparticles: Catalytic labels for the electrochemical detection of α-1-fetoprotein based on TiO2 nanoparticles synthesized with ionic liquid

A novel strategy for the preparation of amperometric immunosensor for rapid determination of α-1-fetoprotein (AFP) in human serum has been developed. TiO₂ nanoparticles (NPs) were prepared by solvothermal reaction using TiCl₄ as raw materials and the mixture of ionic liquids and doubly distilled water as solvent. α-1-fetoprotein antibody (AFP Ab) was mixed with TiO₂ NPs/chitsotan (CHIT) solution and immobilized onto the surface of a glassy carbon electrode. AFP (Ab) functionalized Au NPs were used as catalytic labels for the amperometric detection of AFP by means of the electrocatalyzed reduction of Au NPs to H₂O₂. The electrochemical behavior of the immunosensor was studied. Other experimental conditions such as pH, immunoreactions temperature and time were also studied. The prepared immunosensor offers an excellent amperometric response for AFP ranging from 1.0 to 160.0 ng/mL with a detection limit of 0.1 ng/mL. The result shows that the immunosensor displays rapid response, high sensitivity, good reproducibility and favorable stability.     

基于蛋白A-纳米金-丝网印刷电极的雌二醇免疫传感器

将蛋白A固定在纳米金修饰的丝网印刷电极表面,纳米金大的比表面积以及较高的表面自由能使蛋白A比较牢固地固定在其上;蛋白A分子可以定向结合17β-雌二醇抗体的Fc片段,使抗体在电极表面有序固定化。利用17β-雌二醇和酶标17β-雌二醇的酶免疫竞争反应,研制了快速测定17β-雌二醇的电化学传感器。用循环伏安法和扫描电子显微镜对修饰过程进行了表征。优化测定条件后,传感器呈现了很高的测定灵敏度,电流响应信号与17β-雌二醇浓度在0.1μg/L～20μg/L范围内呈良好的负线性相关,检出限为0.035μg/L(S/N=3)。应用该免疫传感器测定了正常人群尿样中17β-雌二醇含量,测定范围与报道的高效液相色谱法测定结果相一致,样品加标回收率为96%～114%。     

赤潮毒素大田软海绵酸表面等离子共振免疫检测方法研究

结合表面等离子体共振技术与免疫检测技术,研究和建立了一种响应速度快、免标记、低成本新型海洋赤潮毒素大田软海绵酸检测方法.基于SpreetaTM传感器构建了小型表面等离子共振免疫检测系统,采用共价偶联方法在传感器金膜表面修饰大田软海绵酸-牛血清蛋白抗原作为生物敏感膜;测得该方法相对标准偏差为1.51％ (n=12),定量...     

An ultra-sensitive chemiluminescence immunosensor of carcinoembryonic antigen using HRP-functionalized mesoporous silica nanoparticles as labels

A novel chemiluminescence immunosensor using horseradish peroxidase (HRP)-functionalized mesoporous silica nanoparticles (MSN) as labels was developed, which increases the sensitivity of the chemiluminescence immunoassay. The enzyme-functionalized MSN were fabricated by simultaneous coimmobilization of HRP and the carcinoembryonic antigen antibody (anti-CEA) onto the surface of MSN using 3-aminopropyltriethoxysilane (APTES) as the linkage. Because the large surface area of MSN carriers increased the amount of HRP bound per sandwiched immunoreaction, the conjugates provided a much higher signal and increased sensitivity. This is an improvement over the traditional sandwich immunoassay which often has one or two enzyme molecules per antibody. This approach was successfully demonstrated as a simple, cost-effective, specific, and potent method to detect CEA in practical samples. The analysis showed a linear response within the range of 0.1-40 ng/mL (r = 0.9912). The relative standard deviation (RSD) for 11 parallel measurements of 20 ng/mL CEA was 3.9%. The sensitivity of the immunosensor using MSN-HRP-Ab2 as labels was about 10-fold higher than that of traditional labels. These labels for immunosensors may provide many potential applications for detection of different biomolecules.