生物免疫传感器检测迟缓爱德华氏菌研究

迟缓爱德华氏菌(E.tarda)是最为严重的水产动物致病菌之一,准确、即时的检测手段是预防控制该菌传播的关键所在.通过量子点(QDs)标记E.tarda单克隆抗体(Ab),利用生物免疫传感器技术实现E.tarda的快速、特异性检测.结果显示,QDs-Ab的荧光通过加入氧化石墨烯(GO)产生淬灭,构建了捕获目标细菌的探针,GO最适淬灭浓度为60μg/L.细菌捕获探针中加入E.tarda后,能够检测到重新恢复强度的橙色荧光.针对E.tarda设计的生物免疫传感器的特异性,选取灿烂弧菌、溶藻胶弧菌、副溶血弧菌和嗜水气单胞菌作为对照,结果显示,对照组不能明显引起荧光强度的改变,而实验组却能显著提高荧光强度.本研究建立的基于荧光能量共振转移(FRET)的具有高灵敏度和特异性的生物传感器检测方法在细菌的早期诊断中有良好的应用潜质.     

用于室内有毒气体快速检测的便携式CC/SAW电子鼻

开发了一种用于室内空气质量快速检测的便携式电子鼻气体分析仪.该仪器利用毛细管分离柱(CC)将混合气体选择性分离,实现对气体的分类识别,然后借助声表面波(SAW)传感器频率响应测量每种组分对应的浓度,实现对气体的量化.实验针对17种有毒有害气体进行测试,结果表明该系统能对低浓度复杂混合气体进行快速检测,且具有较高的灵敏度,良好的选择性和重复性.这为不同应用场合下的痕量气体检测提供了一种可行的技术.     

水环境重金属铜离子光学检测仪器的设计

针对水质污染中的重金属污染设计了一种自动化检测仪器,能够特异性检测水环境中重金属铜离子.仪器采用光学方法进行检测,具有检测速度快,操作简单,重复性好等优点.仪器能够自动控制水路实现了溶液进样自动化,同时设计了水路闭环检测模块,实现了精确进样以及故障检测.仪器能够自动调节激光器光强适应于不同浓度样品的检测.实际检测结果表明本仪器检出限达到0.006 mg/L,在0.1 mg/L到2.00 mg/L具有良好的线性度,检测准确度小于6.4%,精密度小于4.52%.     

一种新型唾液sIgA电流型纳米免疫生物传感器的制备

通过层层自组装,将硫堇(Thi)和纳米金(GNPs)修饰到Nation修饰的玻碳(GCE)电极表面,利用纳米金单层吸附唾液分泌性免疫球蛋白A(sIgA),最后用辣根过氧化物酶(HRP)封闭电极上的非特异性吸附位点,构建了一种检测唾液sIgA的新型电流型纳米免疫生物传感器.该生物传感器灵敏度高,特异性好,测试方便,检测线性范围为6.5-300mg/L,检出限为3.0mg/L;电流值达到95%稳态时间小于20s.探讨了抗体和底物浓度,pH值和温度,孵育时间,干扰物对传感器的影响.该传感器与ELISA法相关性良好(R=0.98932,P<0.001),可用于唾液sIgA的快速、准确检测,从而判断人体局部免疫状况.     

适用于鱼肉中氯霉素检测的抗体包被金磁纳米微粒修饰安培免疫传感器研究

研制了基于氯霉素抗体包被Fe3O4/Au金磁纳米微粒(GMP)和三乙撑四胺铜(II)(CuL)共固定修饰平面热解石墨电极的安培免疫传感器(PRG|CuL / anti CAP-GMP),用于测定鱼肉中CAP含量.该免疫传感器是利用外加磁场,将anti CAP-GMP吸引到CuL修饰的PRG电极(PRG|CuL)表面制备而成.CuL对H2O2还原具有良好的电催化能力,当该传感器在含CAP样品液中温育后,CAP与电极表面的anti CAP的免疫结合物导致CuL对H2O2的催化还原电流(I)降低,电流下降值(△I)和CAP浓度成正比,可用于CAP定量测定.在25℃的pH=6.5磷酸盐缓冲液(PBS)中温育30 min,该传感器对CAP的检测线性范围为0.6~110 ng/mL,检出下限为0.092 ng/mL(3σ法).应用于鱼肉中CAP检测并与传统的液相色谱法(HPLC)比较,结果一致,其添加回收率在97%~104%之间.该免疫传感器集分离、富集为一体,电极表面可更新,检测灵敏快速,对于水产品中痕量氯霉素分析提供了一种新颖的方法.     

一次性酶免疫传感电极快速检测福氏志贺氏菌的研究

为研究快速检测福氏志贺氏菌的电化学免疫传感电极,将羧基化多壁碳纳米管(MWCNT)与壳聚糖(chitosan)制备成复合物,应用此复合物将辣根过氧化物酶标记的福氏志贺氏菌抗体(HRP-anti-S.flexnefi)上步直接固定在四通道丝网印刷碳电极表面,制成快速检测福氏志贺氏菌的酶免疫传感电极.采用原子力显微镜表征不同修饰电极的表面形态,循环伏安法考察不同电极的电化学特性和监测酶促反应,利用还原峰峰电流的减小来测定福氏志贺氏菌.在优化的测定条件下,免疫电极对福氏志贺氏菌的检测范围为104～1010cfu/mL,检出限为2.3×104du/mL(S/N=3).而且该酶免疫传感电极具有较好的特异性、重现性、稳定性和准确性,该方法还具有快速、简便、易于操作和价格低廉等优点,具有用于福氏志贺氏菌快速筛检的潜力.     

基于核酸识体的电化学免疫传感器用于血小板源性生长因子的检测

该文提出了一种新颖的、高灵敏度的基于核酸识体和银沉积的电化学免疫传感器用于人体血小板源生长因子BB(PDGF-BB)的检测.首先在电极的表面固定抗体,然后与抗原、生物素标记的核酸识体形成免疫夹心复合物,再利用生物素和亲合素的特异性反应把亲合素标记的碱性磷酸酶固定到电极上,然后把得到的产物在底物溶液中进行银沉积,最后利用电化学方法对沉积到电极表面的银进行检测以达到检测PDGF-BB的目的.实验对固定抗体浓度和生物素标记核酸识体的浓度两个条件进行了优化.实验结果表明PDGF-BB的浓度在1～1000 ng/mL的范围具有良好的线性关系,最低检测下限可达0.8 ng/mL.该方法具有高灵敏度和稳定性,在临床上具有广阔的应用前景.     

食物致病菌快速检测系统的设计与实现

设计了食物致病菌快速检测一体化系统。基于抗原-抗体特异性结合的原理,采用TiO2纳米线束为敏感元件设计微生物免疫传感器,以高集成度芯片AD5933设计阻抗检测电路来通过电化学阻抗谱法测量该传感器中TiO2纳米线束的阻抗变化量,实现了致病菌数量的快速检测。以大肠杆菌为例作为实验样本对检测一体化系统的性能进行测试,实验结果表明该检测一体化系统可以快速检测出食物中存在的大肠杆菌,检测周期约为1 h,系统的检测下限为4.5×102cfu/mL,传感器在扫描频率为1 000 Hz～10 000 Hz之间表现出来的重复性较好。     

前列腺特异性抗体电化学免疫传感器研制

制得自组装巯基丁二胺铜(Ⅱ)/纳米金胶/前列腺特异性(PSA)抗体免疫传感器.在pH=5.2的磷酸盐底液中,采用示差脉冲法对PSA进行定量测定.结果显示:该传感器的氧化峰电流减少值与PSA浓度在0.005～0.48mg/L范围内成线性关系,检测下限为2μg/L.在40μg/L PSA浓度下测量BSD=2.9%(n=8),该免疫传感器稳定性和抗干扰性较好.检测血清中PSA结果满意.     

钻杆螺纹复合电磁检测方法与仪器

钻杆螺纹轮廓面复杂且呈现一定锥度,一直缺少快速有效的检测方法与仪器.提出了一种基于自磁化和永磁磁扰动法的钻杆螺纹复合电磁检测方法.分析了探头扫查方式并对实现其动作的机构进行了设计.开发出相应的检测仪器,并对该仪器进行了测试,结果表明:该检测仪器具有便携、易操作、效率高的特征以及良好实用性和广阔的应用前景.     

A PDMS microfluidic impedance immunosensor for E. coli O157:H7 and Staphylococcus aureus detection via antibody-immobilized nanoporous membrane

In this article, a PDMS microfluidic immunosensor integrated with specific antibody immobilized alumina nanoporous membrane was developed for rapid detection of foodborne pathogens Escherichia coli O157:H7 and Staphylococcus aureus with electrochemical impedance spectrum. Firstly, antibodies to the targeted bacteria were covalently immobilized on the nanoporous alumina membranes via self assembled (3-glycidoxypropyl)trimethoxysilane (GPMS) silane. Then, the impedance spectrum was recorded for bacteria detection ranging from 1 Hz to 100 kHz. The maximum impedance amplitude change for these two food pathogens was around 100 Hz. This microfluidic immunosensor based on nanoporous membrane impedance spectrum could achieve rapid bacteria detection within 2 h with a high sensitivity of 10² CFU/ml. Cross-bacteria experiments for E. coli O157:H7 and S. aureus were also explored to testify the specificity. The results showed that impedance amplitude at 100 Hz had a significant reduction in binding of bacteria when the membrane was exposed to non-specific bacteria.     

检测大肠杆菌O157:H7的电化学阻抗谱生物传感器的研究

我们提出了用掺锡的三氧化二铟(ITO)作为工作电极,通过硅烷化固定化技术,将抗大肠杆菌O157:H7单克隆抗体固定在ITO电极表面,利用电化学阻抗谱技术来构建一种新型的免疫传感器.该新型的免疫传感器的检测限为 4×103CFU/mL,检测线性范围为4×103-4×106CFU/mL.实验研究表明,该传感器具有灵敏度较高,检测时间短,操作简单等优点,在临床医学和环境监测中具有应用价值.     

基于混合损失联合调优与多尺度分类相结合的肺结节检测算法

针对CT图像的肺结节自动检测任务中检测灵敏度低及存在大量假阳性的问题,提出了一个基于混合损失的三维全卷积网络与基于注意力的多尺度三维残差网络相结合的肺结节检测方法。首先,基于相似度损失预训练三维全卷积网络,利用该网络筛选难例样本,并基于混合损失将难例与正样本进行联合调优得到候选结节检测网络,用于快速筛选疑似结节;然后,利用基于注意力的多尺度三维残差卷积网络对疑似结节进行分类,从候选结节中精确地分辨出真正结节。在LUN16数据集上,候选结节检测阶段的灵敏度在每个病例的假阳数目为59.1时达到97.18%,检测系统的平均灵敏度为0.880,表明本算法可以提高肺结节检测的灵敏度并有效控制假阳性,在LUNA16数据集上获得了更优的性能。     

An integrated microfluidic system for live bacteria detection from human joint fluid samples by using ethidium monoazide and loop-mediated isothermal amplification

Periprosthetic joint infection (PJI) is one of the severe complications of prosthetic joint replacement. Delayed PJI diagnosis may anchor bacteria in periprosthetic tissues, and removal of the prosthesis might be inevitable. The diagnosis of PJI depends on the identification of microorganisms by standard microbiological cultures or more advanced molecular diagnostic methods for detection of bacterial genes. However, these methods are relatively time-consuming, labor-intensive and not human error-free. Moreover, it is challenging to distinguish live from dead bacteria by using DNA-based molecular diagnostics since bacterial DNA will be remained in the tissue even after the death of the bacteria. In this work, an integrated microfluidic system has been developed to perform the entire molecular diagnostic process for the PJI diagnosis in a single chip. We combined the loop-mediated isothermal amplification (LAMP) with ethidium monoazide (EMA) in an integrated microfluidic system to identify live bacteria with reasonable sensitivity and high specificity. All the diagnostic processes including bacteria isolation, cell lysis, DNA amplification and optical detection can be automatically performed on the integrated microfluidic system by using a compact custom-made control system. The integrated system can accommodate four primers complementary to six regions of the target genes and improve the detection limit by using LAMP. The limit of detection in this multiple EMA-LAMP assay could be as low as 5 fg/reaction (~1 CFU/reaction) when choosing an optimized primer set as we demonstrated in mecA gene detection. Thus, the developed system for PJI diagnosis has great potential to become a point-of-care device.     

Automated immunomagnetic processing and separation of Legionella pneumophila with manual detection by sandwich ELISA and PCR amplification of the ompS gene

The culture-independent and automated detection of bacteria in the environment is a scientific and technological challenge. For detection alone, a number of sensitive methods are known (e.g., PCR, enzyme-linked immunosorbent assay [ELISA], fluorescent in situ hybridization) but a major problem remaining is the enrichment and separation of the bacteria that usually occur at low concentrations. Here, we present an automated capturing and separation system, which can easily be combined with one of the sensitive detection techniques. We have developed a method for enrichment and detection of Legionella pneumophila in liquid media. Concentrated microorganisms were either detected by PCR or by sandwich ELISA. The limit of detection with the immunological assay was about 750 bacteria. Using PCR, the equivalent of about 2000 genomes could be detected. The assays were then transferred to a laboratory prototype for automated processing. It was possible to automatically enrich L. pneumophila by immunomagnetic separation (IMS), and again, the bacteria were detected by sandwich ELISA and PCR amplification of the ompS gene. As a novel aspect, ompS gene was used for the first time as a target for the detection of L. pneumophila on magnetic beads. The aim of this work was to develop an automated procedure and a device for IMS of bacteria. With Legionella as a model organism, we could show that such a novel fully automated system can be an alternative to time-consuming conventional cultivation methods for detecting bacteria or other microorganisms.     

Use of fuzzy-logic-inspired features to improve bacterialrecognition through classifier fusion

Escherichia coli O157:H7 has been found to cause serious health problems. Traditional methods to identify the organism are quite slow, pulsed-held gel electrophoresis (PFGE) images contain "banding pattern" information which can be used to recognize the bacteria. A fuzzy logic rule-based system is used as a guide to find a good feature set for the recognition of E. coli O157:H7. While the fuzzy rule-based system achieved good recognition, the human inspired features used in the rules were incorporated into a multiple neural network fusion approach which gave excellent separation of the target bacteria. The fuzzy integral was utilized in the fusion of neural networks trained with different feature sets to reach an almost perfect classification rate of E. coli O157:H7 PFGE patterns made available for the experiments.     

基于CdSe/ZnS核壳型量子点的高灵敏、快速细菌计数新方法的研究

以乙酰丙酮镉和硬脂酸锌为前驱体,合成了巯基丙酸修饰的CdSe/ZnS核壳型量子点(QDs)。并将其作为荧光探针,以金黄色葡萄球菌(S.aureus)为目标细菌,建立了一种高灵敏的、简单快速的细菌计数新方法,并借助荧光显微镜成功的进行成像探测研究。通过考察量子点浓度、孵育时间等因素的影响,确定了细菌定量检测的最佳条件。在最优化的实验条件下,体系的相对荧光强度随细菌数量的增加而增大。该方法的线性范围为102CFU/mL～106CFU/mL,检测限为102CFU/mL,线性回归方程为Y=427.586X-677.022(R=0.996 49)。本方法有效克服了传统的细菌计数方法存在的缺陷,具有较高的灵敏度和较好的重现性(实际样品检测的RSD=3.6%～8.1%),且操作简单、检测时间短、成本低,有很好的潜在应用价值。     

高精度臭灵丹微量元素含量检测系统设计

为了实现快速、无损、实时检测和分析臭灵丹中微量元素的含量,现对高精度臭灵丹微量元素检测系统进行设计。使用当前方法检测臭灵丹时,无法在保证微量元素活性的条件下提取微量元素。为此,提出一种基于Taguchi的高精度臭灵丹微量元素含量检测系统设计方法。该方法使用日本理学ZSX100型X射线荧光光谱仪采集臭灵丹微量元素光谱,确定微量元素种类,利用超临界萃取技术中的两大技术超临界流体萃取技术和超临界固体萃取技术萃取出臭灵丹中的微量元素,再经过超滤分离法中直线段、曲线段、水平段三个阶段分离出萃取液中微量元素,以ICP-MS法计算出微量元素的含量,达到对臭灵丹中微量元素含量的高精度检测。实验仿真证明,所提方法可以快速、无损、实时检测和分析臭灵丹中微量元素。     

Error guided design of a 3D vision system

We argue that for a method to be useful in practice, it is necessary to perform its sensitivity analysis and to investigate its robustness to measurement errors. We present here a complete sensitivity analysis of the 3D reconstruction method based on projective geometry. We use this sensitivity analysis to best design a system for the inference of the shape of a block of granite from cameras placed at 90 degrees angular separation. The system has been tested on both real and synthetic data