插拔式压电肿瘤标志物微阵列免疫传感器的研制

以AT切型、基频10 MHz的金膜石英晶体作为换能器,通过螺旋检测池固定夹具构建一种新型插拔式压电石英晶体传感器,并组装成2×5型压电肿瘤标志物微阵列免疫传感器。研究了传感器的响应特性及参数。该微阵列传感器在甲胎蛋白(AFP)、癌胚抗原(CEA)、前列腺特异性抗原(PSA)和人绒毛膜促性腺激素(hCG)质量浓度分别为20~640μg/L、1.56~50μg/L、1.25~50μg/L、2.5~250 mIU/mL的范围内,压电石英晶体振荡频率偏移值对肿瘤标志物浓度均呈现良好的响应特性。应用微阵列传感器测定68例临床血清标本,结果与化学发光免疫分析法符合(相关系数分别为0.92、0.90、0.91、0.94)。该压电肿瘤标志物传感器微阵列具有结构简单、操作方便、稳定性好、灵敏度和特异性高,不需标记,能实时检测和重复使用等优点,可用于临床实验诊断,具有临床推广应用价值。     

抗体固载于TiO2多孔膜的压电免疫型细菌传感器

在镀银的压电石英晶体上沉积一层TiO2纳米粒多孔膜,用3 氨丙基三乙氧基甲硅烷将其活化后,借助于戊二醛实现了特异性抗体(抗肠道沙门氏菌抗体)在压电石英晶体上的有效固载,并用于肠道沙门氏菌的快速检测。其检测下限为4×104cells mL,检测时间为30min。可用于价格低廉的镀银石英晶体,有望成为开发一次性压电型免疫检测探头的有效方法。     

压电免疫传感器法检测蓖麻毒素

利用纳米金对石英晶体微天平(QCM)的表面修饰和质量扩增效应,建立了一种压电免疫传感器检测蓖麻毒素的新方法。首先在石英晶体的金电极上依次自组装1,6-己二硫醇和纳米金进行表面修饰,然后通过蛋白A定向固定蓖麻毒素多抗来制备敏感膜。利用纳米金的质量扩增效应设计了一种“毒素-单抗-蛋白A-纳米金”复合物,成功实现了对蓖麻毒素的检测,提高了传感器灵敏度和特异性。该传感器对蓖麻毒素响应的线性范围为0.50~10 mg/L,回归方程为△F=45.81Cric in 48.48(R=0.9986,N=10,P<0.01);检测灵敏度为45.81 Hz/(mg/L)。     

基于分子印迹技术的丙溴磷压电石英晶体微天平研制

介绍了一种用于检测丙溴磷农药的分子印迹压电生物传感器的构建方法。采用沉淀聚合法合成了农药丙溴磷的分子印迹聚合物,将其固定于石英晶体微天平电极表面构建传感器;采用环境扫描电镜以及原子力显微镜对聚合物形貌、传感器电极表面形貌特征进行分析,并利用传感器对丙溴磷农药进行检测分析,其质量浓度在10~1000 ng/mL范围内,传感器频率改变与丙溴磷浓度之间的响应呈线性关系,线性方程为y=0.139ρ+2.26(r=0.9984)。结果表明,构建的分子印迹压电生物传感器能够对农药进行初步检测,具有较高的灵敏性和较好的特异识别能力。     

压电晶体传感器的研究进展

本文简要介绍了压电晶体传感器的基本原理,以及基于质量、粘度、电导率变化的溶液分析法。重点介绍了电化学石英晶体微天平（EQCM）、压电生物传感器;对具有很大发展潜力和重要应用价值的串联式压电传感器（SPQC）、串联式表面声波电导传感器（SAW）、液隔电极式压电传感器（ESPS）等也作了简要说明。     

抗体固载于电聚合物膜的压电免疫型细菌传感器

采用电化学技术在镀银的压电石英晶体上沉积一层聚间苯三酚膜 ,用二乙烯基砜将其活化后 ,实现了特异性抗体 (抗肠道沙门氏菌抗体)在压电石英晶体上的固载 ,并用于肠道沙门氏菌的快速检测 ;其检出限为5×104cell/mL,检测时间为30min;该抗体固载技术简单易行 ,重复性好 ,并可用于价格低廉的镀银石英晶体 ,可望成为开发一次性压电型免疫检测探头的有效方法     

压电胰岛素-C肽微阵列免疫传感器研究

以AT切型、基频10MHz的镀金膜石英晶体作为换能器,将抗人胰岛素和C肽单克隆抗体固定在石英晶体电极表面,用2×5检测池固定夹具构建一种新型压电胰岛素-C肽微阵列免疫传感器.研究了抗体固定方法、抗体工作浓度、固定量、一致性以及传感器的响应参数如检测温度、时间和特异性等的影响.该微阵列传感器在胰岛素浓度为2.5～160.0mIU/L、C肽浓度为0.375～12.0ng/mL范围内响应特性良好,压电晶体频率偏移值与胰岛素和C肽浓度呈良好的线性关系.将此微阵列传感器用于人血清标本的测定,结果与放射免疫法符合(r为0.92和0.94).此微阵列传感器具有灵敏度高、特异性好,低密度阵列结构,检测通量较高,不需标记,操作简单、能实时在线检测和重复使用等优点,能用于临床实验诊断,具有临床推广应用价值.     

压电质量传感及其应用

本文介绍了压电质量传感即石英晶体微天平（ＱＣＭ）方法的原理及应用。深入探讨了非质量因素对晶振频率的影响、传感器响应讯号的测定方法。综述了压电免疫传感器的研制概况。提出了今后研究工作的方向。     

基于酶催化银沉积质量放大的血吸虫压电免疫传感器的研究

发展了一种基于酶催化金属银沉积信号放大的新型高灵敏气相压电免疫传感检测技术.先将血吸虫抗原(SjAg)共价固定在石英晶体表面,制备得到血吸虫压电免疫传感器.检测时,在晶振上滴加不同浓度的待测血吸虫抗体,再将碱性磷酸酶标记的二抗通过夹心方式结合到传感器表面.然后利用碱性磷酸酶催化磷酸化的抗坏血酸酯水解从而还原硝酸银,使金属银沉积在晶振表面上,放大传感器的质量响应信号.实验结果表明该传感检测方法可显著提高气相压电免疫传感器的检测灵敏度,传感器对血吸虫抗体的响应线性范围在1～225 ng/mL,检测下限为1 ng/mL.     

新型压电肿瘤标志物微阵列免疫传感器的研究

采用螺旋固定夹具构建一种新型插拔式压电石英晶体传感器,巯基化自组装技术固定抗人甲胎蛋白(AFP)、癌胚抗原(CEA)和前列腺特异抗原(PSA)的单克隆抗体,并组装成2×5型压电肿瘤标志物微阵列免疫传感器.研究了压电肿瘤标志物微阵列免疫传感器的响应特性及其影响因素.该微阵列传感器在AFP、CEA和PSA浓度分别为20～640 μg/L、1.56～50 μg/L和1.25～50 μg/L,压电石英晶体振荡频率偏移值对肿瘤标志物浓度呈现良好的响应特性.微阵列传感器用于68例临床血清标本的测定,结果与免疫化学发光法一致;相关系数分别为0.92、0.90和0.91.     

A Novel QCM-based Biosensor for Detection of Microorganisms Producing Hydrogen Sulfide

Abstract

A novel piezoelectric quartz crystal microbalance (QCM) device with gas permeable membrane is proposed for the detection of microorganisms producing hydrogen sulfide (H₂S). The detection theory is based on the adsorption of hydrogen sulfide onto the silver electrode of the piezoelectric crystal sensor, which causes a dramatic decrease in the resonant frequency of QCM. A 100 Hz frequency shift is chosen as the criteria value to judge the presence of microorganisms producing H₂S. Factors affecting detection were investigated. Desiccant is of great practical importance in sensor response. This new biosensor can be a potential candidate for detecting bacteria which produce hydrogen sulfide.     

A coated piezoelectric crystal detector for the selective detection and determination of hydrogen sulfide in the atmosphere

A method for the selective detection and determination of hydrogen sulfide in the atmosphere is presented. This method utilizes the reversible adsorption of H₂S on a piezoelectric quartz crystal coated with an acetone extract of soots resulting from the burning of organochlorine compounds. The extract of a soot prepared from chlorobenzoic acid provided the best substrate. The method is useful in the concentration range 1–60 ppm.     

Development of an Amperometric Microbial Biosensor Based on Thiobacillus thioparus Cells for Sulfide and Its Application to Detection of Sulfate‐Reducing Bacteria

This paper describes a novel electrochemical microbial biosensor based on Thiobacillus thioparus cells for sulfide detection. The morphology and electrochemical properties of the proposed biosensor were characterized by SEM and cyclic voltammetry, respectively. Working conditions of the microbial biosensor were optimized to obtain good electrochemical performances. Under the optimum conditions, analytical performances were evaluated, and the results suggested that the microbial biosensor could be used for selective detection of sulfide. The microbial biosensor was then successfully applied in detection of sulfate‐reducing bacteria by oxidizing its characteristic metabolite, sulfide, which was accumulated in culture media during bacterial growth.     

A coated piezoelectric crystal detector for the determination of hydrogen sulfide

A method is presented for the detection and determination of hydrogen sulfide. It is based on the selective and reversible adsorption of hydrogen sulfide on the surface of a piezoelectric crystal coated with a cadmium iodide/urea/-glycerol solution. The proposed detector has a response time of 30 s with linear response over the concentration range 2–25 μl l^?1 (ppm).     

QCM-DNA biosensor based on plasma modified PT/TiO2 nanocomposites

A quartz crystal microbalance DNA biosensor based on plasma prepared polythiophene /titanium dioxide (PT/TiO₂) nanocomposite was developed for the detection of genetically modified organisms (GMOs). DNA hybridization was studied by quartz crystal microbalance (QCM) and cyclic voltammetry (CV) measurements. Single stranded DNA probes were immobilized on the PT/TiO₂ coated quartz crystal electrode and the hybridization between the immobilized probe and the target complementary sequence in solution was monitored. The developed QCM-DNA biosensor represented promising results for a real-time, label-free, direct detection of DNA samples for the screening of genetically modified organisms.     

High sensitivity microgravimetric biosensor for qualitative and quantitative diagnostic detection of polychlorinated dibenzo-p-dioxins

A piezoelectric immunosensor system was developed for the rapid detection of polychlorinated dibenzo-p-dioxins (PCDDs). The system uses a competitive inhibition enzyme immunoassay (EIA) based on a mouse monoclonal antibody that is specific for 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and a conjugate of a dioxin-like competitor coupled to the enzyme horseradish peroxidase (HRP). The anti-dioxin antibody was deposited on a 10 MHz AT-cut quartz crystal resonator modified with a self-assembly monolayer of dithiobis-N-succinimidyl propionate. PCDDs at different concentrations in the range 0.001-10 ng mL-1 were mixed with a constant amount of HRP-conjugated competitor. The frequency responses due to the adsorption of the mixed samples on the biosensor surface were measured. The results show that 2,3,7,8-TCDD can be quantitatively detected with the developed immunosensor in the concentration range 0.01-1.3 ng mL-1. Cross-reactivities of the biosensor to various PCDD congeners were also investigated. The sensitivity and selectivity of the quartz crystal microbalance (QCM) biosensor is comparable to EIA and ELISA methods in the detection of polychlorinated dibenzo-p-dioxins. The developed QCM immunosensing system offers significant improvements in speed, sample throughout and cost for the qualitative and quantitative detection of PCDDs compared with GC-MS.     

A new piezoelectric response model for population growth of bacteria

A piezoelectric response model on the population growth of microorganism is proposed. This model is based on a novel population growth model, which has a more obvious ecological meaning and the fact that the series piezoelectric quartz crystal (SPQC) sensor responses to conductivity changes of the medium during the growth of the microorganism. From the response model four parameters can be obtained including the maximum specific growth rate mu(m), saturated population size N(m), and two constants C and K(1). The influence of the parameters on the response curve is discussed in which the influences of mu(m) and N(m) are more obvious. With the proposed model the quantitative determination of bacteria may be more accurate than the frequency detection time (FDT) method. Then the growth of Escherichia coliform (E. coli) monitored with a SPQC sensor is compared with the simulated growth curve obtained by the proposed model and a good agreement is obtained.     

Detection of fragmented genomic DNA by PCR-free piezoelectric sensing using a denaturation approach

Label-free and real-time DNA sequence detection in PCR-amplified DNA samples can now be achieved by different approaches. On the contrary, only few works have been reported dealing with direct sequence detection in nonamplified genomic DNA. Here, a piezoelectric biosensor for direct detection of sequences in nonamplified genomic DNA is described. The system relies on real-time and label-free detection of the hybridization reaction between an immobilized probe and the complementary sequence in solution. The DNA probe is immobilized on the sensing surface (10 MHz quartz crystals), while the complementary sequence is present in the genomic DNA, previously fragmented with restriction enzymes.     

A Piezoelectric Biosensor For DNA Hybridisation Detection

Abstract

In this paper the realisation of a piezoelectric biosensor for DNA hybridisation detection is reported. A biotinylated 25-mer, was immobilised on a (strept)avidin coated piezoelectric crystal; avidin was covalently bound to the thiol/dextran modified gold surface of the crystal. Hybridisation of the probe with a complementary sequence was detected. The device was able to distinguish among targets of different lengths. Many cycles of measurements could be performed on the same crystal surface regenerating the single strand with HCl (ImM). No signal was detected when the probe reacted with a non complementary sequence. The same experiments were performed immobilising the biotinylated DNA probe on the gold surface coated with avidin by adsorption and the results were compared.     

Detection of β-thalassemia by a DNA piezoelectric biosensor coupled with polymerase chain reaction

β-Thalassemia is an inherited disorder mainly caused by mutations in the gene of the β-globin chain of adult haemoglobin (HbA). Clinically, β-thalassemia can be a mild or silent condition, or it can cause severe diseases, leading to transfusion dependence. Studies at the gene level have identified a large number of variations in the β-globin gene in different populations. In the Mediterranean area one of the most common mutation is the C → T substitution in the codon 39 of the gene.A new procedure for detecting codon 39 mutation in the β-globin gene is reported, based on a DNA piezoelectric biosensor. An oligonucletidic probe (25-mer), specific for the region around codon 39, is immobilised on the gold surface of a piezoelectric quartz crystal. The hybridisation between the immobilised probe and the complementary strand in solution is detected recording the variations of the crystal frequency.Experiments with synthetic oligonucleotides were initially performed. Distinguishable frequency shifts were obtained from the interaction between the immobilised probe and the complementary and the mismatch oligonucleotides. A solution containing 50% of both the oligonucleotides has been also tested and distinguished from the others evaluating the resulting signals. Experiments with non-complementary oligonucleotides gave no signal variation. The biosensor was able to distinguish between sequences differing in only one base also using polymerase chain reaction-amplified samples [771 base pairs (bp)] of DNA extracted from human blood of thalassemic and healthy (normal) patients or patients with β-thalassemia traits.The optimised DNA piezoelectric biosensor has been successfully applied to the determination of one of the most frequent mutation characteristic of β-thalassemia in the Mediterranean population.