纳米金/氧化石墨烯—离子液体修饰电极测定多菌灵

制备带正电的氧化石墨烯—离子液体(GO-IL)复合材料,用带负电的纳米金(GNP)自主装到GO-IL表面,构建GNP/GO-IL/GC修饰电极。利用电化学阻抗谱对所制备的修饰电极进行表征,在0.2～10μmol/L浓度范围内,多菌灵的差分脉冲伏安峰电流与浓度呈现良好的线性关系,相关系数R为0.997,检出限为0.08μmol/L。并用差分脉冲伏安法对砂糖橘中多菌灵含量进行了测定,结果表明,该方法简便、快捷、灵敏度高。     

基于表面增强拉曼光谱适配体传感器检测植物蛋白肉中的黄曲霉毒素B1

目的 随着生活水平的提高,经济社会的发展,面对丰富多样的食品种类,保证饮食安全已成为民众关注的重点。然而,食品中的黄曲霉毒素B1（Aflatoxin B1,AFB1）的存在对人类健康构成了极大的威胁。因此,开发一种快速、简单、灵敏的AFB1检测方法,对食品安全十分必要和迫切。方法 研发了一种检测AFB1的表面增强拉曼散射(surface enhancement of Raman scattering, SERS)适配体传感器,传感器由巯基适配体（SH-DNA）修饰的金包四氧化三铁磁性纳米粒子（Fe3O4@AuMNPs）作为捕获底物与巯基适配体互补链（SH-cDNA）修饰的金包二氧化硅纳米粒子（SiO2@Au-MBANPS）作为信号探针所组成。通过适配体与AFB1的特异性结合可以导致捕获底物释放信号探针,SERS信号强度随着AFB1的浓度的变化而发生变化,从而实现对AFB1的检测。结果 通过实验条件的优化,得到线性回归方程为Y= -222.07lgX+ 5723.12,r2= 0.9986的标准曲线,检出限为0.37 pg/mL。该SERS传感器成功应用于实际样品中AFB1的分析,得到回收率为98% ~ 110%,相对标准偏差值为3.9% ~ 5.7%。结论 本研究建立的SERS适配体传感系统可以实现对AFB1的定量检测,可以清楚识别AFB1与其他毒素的SERS信号强度差别,为AFB1的快速检测提供了新的检测方法,在实际样品植物蛋白肉中表现出优异的检测性能。     

基于核酸适体-纳米金银染放大的玉米赤霉烯酮快速检测方法研究（网络首发、推荐阅读）

玉米赤霉烯酮（Zearalenone,ZEA）具有较强的生殖毒性、致突变和致畸作用。以ZEA适体为识别元件,构建了基于纳米金诱导聚集和银染放大的ZEA适体比色可视化检测方法。结果表明,在优化条件下,ZEA浓度在5~200 ng/mL范围内与体系的吸光度值呈良好的线性关系,其线性回归方程为y= 0.248 6+0.000 461 56x (R2=0.990 2),最低检测限为5 ng/mL,且方法特异性良好。进一步通过银染作用将该方法的灵敏度提高了50倍。经对比,该方法对实际样品的检测结果与酶联免疫法基本一致,为食品中ZEA的快速检测提供了简便有效的新策略。     

Sensitive enzyme‐linked immunosorbent assay and rapid one‐step immunochromatographic strip for fumonisin B1 in grain‐based food and feed samples

BACKGROUND: Maize contaminated with mycotoxin fumonisin B1 poses a global threat to agricultural production. In this study, polyclonal antibodies (pAb) specific to fumonisin B1 were generated from rabbits immunised with fumonisin B1–keyhole limpet haemocyanin (KLH). These antibodies were used to establish a sensitive competitive direct enzyme‐linked immunosorbent assay (cdELISA) and gold nanoparticle immunochromatographic strip for detecting fumonisin B1 levels in maize‐based foods and feeds. RESULTS: In cdELISA, fumonisins B1, B2 and B3 at concentrations of 0.42, 0.58 and 81.5 ng mL^?1 respectively caused 50% inhibition (IC₅₀) of binding of fumonisin B1–horseradish peroxidase (HRP) to the antibodies. Effective on‐site detection of fumonisin B1 was achieved by developing a rapid and sensitive pAb‐based gold nanoparticle immunochromatographic strip. This strip had a detection limit of 5 ng mL^?1 for fumonisin B1 in maize‐based samples. Additionally, the whole analytical process could be completed within 10 min. Close examination of 15 maize‐based samples by cdELISA revealed that 11 were fumonisin‐positive, with a mean concentration of 435 ± 20.1 ng g^?1. These results correlated well with those obtained by immunochromatographic strip. CONCLUSION: Both cdELISA and immunochromatographic strip methods established in this study are sensitive for rapid detection of fumonisins in agricultural commodities. Copyright © 2010 Society of Chemical Industry     

Voltammetric detection of Cr(VI) with disposable screen-printed electrode modified with gold nanoparticles

Gold nanoparticle (Au-NP) enhanced voltammetric detection of Cr(VI) is developed for determination of trace amounts of Cr(VI) in an acetate buffer media (pH 4.6). The Au-NPs were electrodeposited onto a disposable screen printed electrode (SPE) via an electrodeposition step. It was found that the electrodeposited Au-NP has strong adsorption on Cr(VI) species, which results in an enhanced reduction current of Cr(VI). Compared with the bulk gold electrode, the reduction current of Cr(VI) was enhanced 10 times with the Au-NP-modified SPE electrode. Square wave voltammetric (SWV) measurement with the disposable Au-NP-modified SPE provides a fast, simple and sensitive detection of trace amounts of Cr(VI). The adsorption of Cr(VI) on Au-NP was characterized with voltammetry, X-ray photoelectron spectroscopy and ultraviolet spectra. The different parameters including the electrodepositing time, supporting electrolyte, and pH that govern the analytical performance of the electrode have been studied in detail and optimized. The detection limit of 5 microg L(-1) Cr (VI) was obtained under optimum experimental conditions. The performance of the sensor was successfully evaluated with river water samples spiked with Cr(VI), indicating this convenient and sensitive technique offers great promise for onsite environmental monitoring and biomonitoring of Cr(VI).     

Colloidal gold-based immunochromatographic test strip for rapid detection of abrin in food samples

In the present study, we developed a convenient, rapid, and sensitive immunochromatographic (IC) test strip to detect abrin in assay buffer and spiked abrin in test food samples. The abrin IC test strip was based on a sandwich format consisting of a monoclonal antibody and a polyclonal antibody. The anti-abrin A chain monoclonal antibody from mice was immobilized on a porous nitrocellulose membrane as a capture antibody, while the anti-abrin polyclonal antibody from rabbits was conjugated to colloidal gold particles, serving as a detection antibody. Both visual observation and quantitative analysis indicated that the lower detection of the strip was about 3 ng/ml when abrin was directly spiked into milk, orange juice, and drinking water at a concentration of 3 to 60 ng/ml; the analytical recovery rate was 92.2 to 128%. With this method, abrin spiked into food could be detected in less than 10 min. Moreover, the IC test strip showed no cross-reaction with the closely related phytotoxin ricin. Therefore, our test strip is an ideal candidate for the development of a kit for rapid and quantitative detection of abrin in food samples.     

酰肼介导的抗体定向偶联策略提高免疫层析试纸条检测性能

以酰肼功能团修饰的胶体金作为试纸条的标记材料,利用酰肼基团可特异性地与抗体Fc片段醛基发生亲核加成反应的特性,实现抗体在胶体金纳米粒子（gold nanoparticles,AuNPs）表面的定向偶联。对比于碳二亚胺介导的共价偶联法,酰肼功能团介导的定向偶联需要更少的抗体标记量,更短的偶联时间,酰肼化胶体金免疫层析试纸条（hydrazided gold nanoparticles-based immunochromatographic assay,hAuNPs-ICA）检测玉米赤霉烯酮（zearalenone,ZEN）展示出更好的检测灵敏度;将hAuNPs-ICA检测ZEN加标的玉米样品,结果显示加标回收率介于83.1%～118.0%,批内、批间变异系数为3.9%～13.1%,表明hAuNPs-ICA具有较好的精密度和准确性。     

Preparation of scFv-immobilized quartz crystal microbalance sensor by PS-tag-mediated solid-phase refolding

The activation of a single-chain Fv antibody on the surface of a quartz crystal microbalance (QCM) sensor chip was investigated in order to develop an economical and sensitive immuno-QCM sensor system for use in clinical diagnosis. On the bare gold surface of a QCM sensor chip, approximately 60nm of hydrophilic polystyrene (phi-PS) thin film that specifically binds the polystyrene-binding peptide (PS-tag) was prepared by spin-coating and O(2)-plasma irradiation. When the adsorption of PS-tag-fused anti-ED-B scFv (scFv-PS) onto the phi-PS surface was directly monitored, the maximum density of scFv-PS attained was 1.56μg/cm(2), 1.6-times higher than that of scFv. The specific antigen-binding activity of scFv-PS after solid-phase refolding increased with the density of immobilized scFv-PS, and, consequently, activity 1.7 times higher than that of scFv was retained. The scFv-PS-immobilized QCM sensor chip rapidly allowed the detection of clear signals for antigen at the range of 0.1-10μg/ml, while no signal was detectable for 10μg/ml BSA as a negative control. The scFv-PS-immobilized QCM sensor developed in the present study will therefore be very useful for the rapid and highly sensitive detection of biomarkers, and should be applicable to clinical diagnosis.     

Sensitive enzyme-linked immunosorbent assay and gold nanoparticle immunochromatocgraphic strip for rapid detecting chloramphenicol in food

Antibody specific to chloramphenicol (CAP) was produced from rabbit that had been immunized with CAP-keyhole limpet hemocyanin (KLH). Using the antibodies, we established a sensitive direct competitive enzyme-linked immunosorbent assay (dcELISA) and a gold nanoparticle immunochromatographic strip (immunostrip) for detection CAP in food samples. In the dcELISA, CAP at levels of 0.15 ng/ml causes 50% inhibition (IC₅₀) of the binding of CAP-horseradish peroxidase to the antibodies. The overall analytical recoveries of CAP (0.25–100 ng/g) added to the honey or milk samples in the dcELISA were 81.9 and 73.7%, respectively. Onsite determination of CAP was accomplished by immunostrips with a detection limit of 0.5 ng/ml and completed within 10 min. Carefully studying 10 honey and 6 milk samples using the dcELISA and immunostrip indicated that all examined samples were negative for CAP. The presented dcELISA and immunostrip methods are sensitive enough for the rapid determination of CAP in the samples.     

Bacterial pathogen detection by conventional culture‐based and recent alternative (polymerase chain reaction,isothermal amplification,enzyme linked immunosorbent assay,bacteriophage amplification,and gold nanoparticle aggregation) methods in food samples: A review

The rapid, sensitive, and selective detection of foodborne pathogens is important to ensure food safety. Culture medium‐based methods for bacteria detection have long been used since Robert Koch's first finding. These methods are simple and cheap but have limitations, such as being time‐consuming, labor‐intensive, and having low selectivity. In this regard, several alternative detection methods have been reported. Among these, recent studies related to the application of polymerase chain reaction, isothermal amplification, bacteriophage amplification, enzyme‐linked immunosorbent assay, and gold nanoparticle aggregation for detection of pathogens in food are discussed in this review. The principles, advantages, and disadvantages of alternative methods are covered, including their rapidity, sensitivity, and selectivity. Finally, regulations related to bacterial pathogen detection in the United States and South Korea were compared with remarks for their progress.     

Modified paramagnetic beads in a microfluidic system for the determination of zearalenone in feedstuffs samples

In this work, we have developed and characterised a novel microfluidic immunoassay methodology for rapid and sensitive quantification of ZEA in feedstuffs samples. The detection of ZEA was carried out using a competitive direct immunoassay method based on the use of anti-ZEA monoclonal antibodies immobilized on magnetic microspheres 3-aminopropyl-modified manipulated for an external remobilize magnets. The ZEA in feedstuffs sample is allowed to compete with ZEA-horseradish peroxidase (HPR) conjugated for the immobilized anti-ZEA antibody. The HPR, in the presence of hydrogen peroxide (H₂O₂) catalyses the oxidation of 4-tert-butylcatechol (4-TBC) whose back electrochemical reduction was detected on gold electrode at 0.0 V. The calculated detection limits for electrochemical detection and ELISA procedure were 0.41 and 2.56 μg kg⁻¹ respectively, the intra and inter-assay coefficients of variation were below 6.5% and the total assay time was 30 min. The microfluidic immunosensor showed higher sensitivity and lower detection limits than the standard ELISA method, which shows potential for detecting ZEA in foods and feeds diagnosis.     

金纳米粒子修饰电极循环伏安法测定食品中的碘

为寻求一种快速、简便、灵敏的食品中碘的测定方法,利用循环伏安法（CV）构建金纳米粒子修饰电极检测碘离子（I-）体系。利用甲烷氧化菌素（Mb）原位还原纳米金（Mb@AuNPs）,电沉积法制备自组装修饰电极。通过透射电子显微镜对Mb@AuNPs表征,CV考察碘离子的电化学行为。确定碘离子检测的优化条件为:电沉积扫描速率0.11 V/s、扫描圈数30圈、缓冲溶液浓度0.05 mol/L、缓冲溶液pH6.5。氧化峰电流与I-浓度在0.01~10.00 μmol/L范围内有良好的线性关系,R2为0.9992,检出限为2.88 nmol/L（S/N）,定量限为9.60 nmol/L,该方法检测不同食品中碘含量的加标回收率为96.22%~103.57%。结果表明该修饰电极对I-的测定具有良好的精密度、稳定性和重现性,以及较好的抗干扰能力,符合测定方法要求,可用于实际样品中碘的测定。     

Underpotential deposition-anodic stripping voltammetric detection of copper at gold nanoparticle-modified ultramicroelectrode arrays

The sensitive detection of copper (II) at gold nanoparticle-modified ultramicroelectrode arrays (UMEAs) is reported. Gold nanoparticles were electrodeposited onto the UMEAs surface by applying a constant positive potential of 1.6 V for 20 min in a 20-nm gold nanoparticle solution. This process significantly increases the electrode area without losing the UMEAs analytical features. Underpotential deposition-anodic stripping voltammetry of copper (II) with such modified UMEAs was performed and showed a high increase in sensitivity (25.9 +/- 1.3 nC x micro-1) and a broader linear range of response (0-10 microM) compared with those values obtained using bare UMEAs (7.5 +/- 0.6 nC x microM(-1) and 0-2 microM, respectively). The copper content of acid extracts of contaminated soils was successfully determined with the modified UMEAs and results are in good agreement with those obtained using the ICP-AES standard method. Overall, this work shows an alternative easy-to-use novel miniaturized device for the rapid and reliable determination of copper in soil samples whose application could be readily extended to other heavy metals of environmental interest.     

Label-free chemiresistive immunosensors for viruses

We report development, characterization, and testing of chemiresistive immunosensors based on single polypyrrole (Ppy) nanowire for highly sensitive, specific, label free, and direct detection of viruses. Bacteriophages T7 and MS2 were used as safe models for viruses for demonstration. Ppy nanowires were electrochemically polymerized into alumina template, and single nanowire based devices were assembled on a pair of gold electrodes by ac dielectrophoretic alignment and anchored using maskless electrodeposition. Anti-T7 or anti-MS2 antibodies were immobilized on single Ppy nanowire using EDC-NHS chemistry to fabricate nanobiosensor for the detection of corresponding bacteriophage. The biosensors showed excellent sensitivity with a lower detection limit of 10(-3) plaque forming unit (PFU) in 10 mM phosphate buffer, wide dynamic range and excellent selectivity. The immunosensors were successfully applied for the detection of phages in spiked untreated urban runoff water samples. The results show the potential of these sensors in health care, environmental monitoring, food safety and homeland security for sensitive, specific, rapid, and affordable detection of bioagents/pathogens.     

Rapid detection of picloram in agricultural field samples using a disposable immunomembrane-based electrochemical sensor

Picloram, a widely used chlorinated herbicide, is quite persistent and mobile in soil and water with adverse health and environmental effects. It is essential to establish a rapid and sensitive method for accurate detection of trace picloram in agricultural samples. We employed a disposable, nontoxic, and conductive chitosan/gold nanoparticles composite membrane on electrochemical sensor for the sensitive detection of picloram in several agricultural field samples. A self-synthesized picloram antibody was encapsulated in the immunomembrane to form an immunoconjugate by a competitive immunoreaction in sample solution, followed by the immobilization of horseradish peroxidase (HRP)-labeled secondary antibody. The immunomembrane possessed good reproducibility for fabrication in batch, providing a congenial microenvironment for the immune molecules. The diffused colloidal Au nanoparticles shuttled the electron transfer between the immobilized HRP and the electrode surface. To demonstrate the suitability of the immunosensor for on-site detection, rice, lettuce, and paddy field water were spiked with picloram and assayed without preconcentration. Under optimal conditions, picloram could be detected in the range from 0.005 to 10 microg/mL with the correlation coefficient of 0.9937, and the detection limit is 5 ng/ mL. The proposed immunosensor exhibited good precision, sensitivity, selectivity, and storage stability.     

Multiplexed detection of mycotoxins in foods with a regenerable array

The occurrence of different mycotoxins in cereal products calls for the development of a rapid, sensitive, and reliable detection method that is capable of analyzing samples for multiple toxins simultaneously. In this study, we report the development and application of a multiplexed competitive assay for the simultaneous detection of ochratoxin A (OTA) and deoxynivalenol (DON) in spiked barley, cornmeal, and wheat, as well as in naturally contaminated maize samples. Fluoroimmunoassays were performed with the Naval Research Laboratory array biosensor, by both a manual and an automated version of the system. This system employs evanescent-wave fluorescence excitation to probe binding events as they occur on the surface of a waveguide. Methanolic extracts of the samples were diluted threefold with buffer containing a mixture of fluorescent antibodies and were then passed over the arrays of mycotoxins immobilized on a waveguide. Fluorescent signals of the surface-bound antibody-antigen complexes decreased with increasing concentrations of free mycotoxins in the extract. After sample analysis was completed, surfaces were regenerated with 6 M guanidine hydrochloride in 50 mM glycine, pH 2.0. The limits of detection determined by the manual biosensor system were as follows: 1, 180, and 65 ng/g for DON and 1, 60, and 85 ng/g for OTA in cornmeal, wheat, and barley, respectively. The limits of detection in cornmeal determined with the automated array biosensor were 15 and 150 ng/g for OTA and DON, respectively.     

生物传感法检测食品中的触酶阳性菌

目的：建立一种检测食品中触酶阳性菌的生物传感法。方法：在一次性丝网印刷电极上聚合辣根过氧化物酶制备检测食品中触酶阳性菌的生物酶传感器,将丝网印刷电极插入电极插口与电化学工作站相连,组装成检测触酶阳性菌的生物电化学传感仪,采用时间- 电流法记录响应的结果。结果：对不同来源的食品,当触酶阳性菌菌落数达到104CFU/mL,即可检测到电流变化,检测所需时间根据样品中菌落数的不同而定,一般仅需1～3h即可推测出样品中原有触酶阳性菌含量,最长通过7h 的增菌培养即可推测出含有0.1CFU/mL 的样品。结论：用该方法检测食品中触酶阳性菌便捷、灵敏、准确,可发展成为一种快速检测食品中触酶阳性菌生物传感器。     

Colorimetric Sensing of Tetracyclines in Milk Based on the Assembly of Cationic Conjugated Polymer-Aggregated Gold Nanoparticles

We have developed an unlabeled sensing strategy for tetracycline (TET) detection in aqueous solutions and milk by using an aptamer–gold nanoparticle (AuNP)-based colorimetric method. Aptamers were found to be absorbed on the Au surface which stabilizes the AuNPs against conditions of high poly(diallyldimethylammonium) strength. The introduction of TET had reduced aptamers to form a TET–aptamer complex, so that the following cationic polymers could aggregate AuNPs and cause a remarkable change in color. Through this phenomenon, TET can be detected qualitatively and quantitatively by means of absorbance. This unlabeled strategy saves both the time and cost for detection, which could selectively detect TET at concentrations as low as 1 μM and 45.8 nM for the naked eyes and colorimetric detector, respectively. In addition, this biosensor exhibited high selectivity over other antibiotics with an excellent recovery in the detection of milk. Because of the advantages of this sensor, we hope this will be the basis for an assay for the sensitive, colorimetric detection of a wide range of molecular analytes.     

Cloth-based hybridization array system for expanded identification of the animal species origin of derived materials in feeds

A cloth-based hybridization array system (CHAS) previously developed for the detection of animal species for which prohibited materials have been specified (cattle, sheep, goat, elk, and deer) has been expanded to include the detection of animal species for which there are no prohibitions (pig and horse) in Canadian and American animal feeds. Animal species were identified by amplification of mitochondrial DNA sequences by PCR and subsequent hybridization of the amplicons with an array of species-specific oligonucleotide capture probes immobilized on a polyester cloth support, followed by an immunoenzymatic assay of the bound PCR products. The CHAS permitted sensitive and specific detection of meat meals from different animal species blended in a grain-based feed and should provide a useful adjunct to microscopic examination for the identification of prohibited materials in animal feeds.     

Gold nanoparticles: From synthesis,properties to their potential application as colorimetric sensors in food safety screening

BackgroundFood safety management and guarantee are the basic requirement during food processing, circulation, storage, and marketing. Elevating the ability to evaluate food quality and safety in a rapid, sensitive and reliable manner is of great importance in food industry. Recently, gold nanoparticles due to unique optical property, ease of functionalization and preparation, and high selectivity and sensitivity have received considerable attention in the field of food safety.Scope and approachGold nanoparticles exhibit distinguishable optical characteristic in different aggregated states and thus have been developed into simple colorimetric sensors for the quick detection of chemical contaminants in food samples. Herein, we reviewed the current methods for synthesis and functionalization of gold nanoparticles, strategies for fabrication of gold nanoparticle based colorimetric sensors and their applications in rapid analysis of food contaminant. Moreover, the inherent optical property of gold nanoparticles and their detecting principle have been highlighted. Finally, the main challenges and future efforts in developing such colorimetric sensors for food contaminants detection were discussed.Key findings and conclusionsGold nanoparticles as smart colorimetric sensors conform to the requirement of modern analysis, such as high selectivity, sensitivity, simplicity, celerity, and portability. Thus, they have great potential to be applied as power sensing tools for food safety screening.