基于电化学生物传感器的核酸肿瘤标志物检测研究进展

肿瘤生物标志物可以实现对癌症的早期诊断,监测肿瘤的发展和预后.随着高通量测序技术的发展,各种类型的核酸被发现与肿瘤的发生发展相关,可作为核酸肿瘤标志物,而且它们对癌症的早期诊断有着重要意义.由于核酸大都具有不稳定性、表达的动态性和低丰度等特点,因此快速灵敏并特异地检测该类肿瘤标志物就显得尤为重要.电化学生物传感器作为一类快速发展的检测方法能够满足这些需求.与传统的检测方法相比,电化学分析法彰显高效、灵敏、简便等优点,而且电化学检测设备装置轻便、廉价且易于微型化和集成化.近年来,用于检测核酸的电化学生物传感器取得了相当大的发展,在精准医疗中具有潜在的应用前景.本文从各种类型的核酸肿瘤标志物入手,介绍检测核酸标志物的电化学传感器在近年来的发展情况,特别是对电化学传感器体系中的探针设计和信号放大策略两个方面进行重点介绍.     

微流控技术在细胞转染中的应用

微流控技术是在微米级、纳米级结构中操控纳升至皮升体积流体的技术与科学,具有液体流动可控、消耗试样和试剂极少等优点。近年来,细胞转染技术有逐渐向微型化技术途径发展的趋势,这也给了研究者从微尺度角度审视细胞转染技术过程的新机会。以下介绍了基于微流控技术的细胞转染方法,包括微阵列方式的转染技术、缩微流动空间中的转染、微流控液滴技术应用于细胞转染、微流控注射技术以及微流控电穿孔技术,并阐述了影响转染效率的因素或改善途径。     

微流控器件-质谱联用接口技术研究进展与应用

生物分析是生命科学研究中的重要环节,分析仪器的小型化是提高生物分析灵敏度、速度、通量和降低成本的有效途径之一.微流控技术能够方便地操纵微量样品,具有集成度高、样品耗量小、污染少等诸多其他常量流控技术难以具备的优点,适用于进行多通道样品处理和高通量分析.除广泛采用的光学和电化学检测手段外,质谱也被用作这些微流控器件的检测器,并逐渐形成了微流控器件-质谱联用技术专门研究领域,进一步促进了自动化程度好、灵敏度高、特异性强的高通量生物分析方法的迅速发展.在大量调研国内外文献的基础上,对微流控器件-质谱联用领域的研究背景和现状进行了综述,不但介绍了微流控器件的制造技术还着重介绍了微流控器件-质谱联用技术在蛋白质组学等生物质谱分析方面的应用和新近进展,评述了可能的发展趋势.     

输血前检测技术的应用与展望

精准输血对疾病治疗、急诊抢救意义重大。开展输血前多项标志物检测是保障精准治疗的关键,是降低输血风险的前提。输血前检测主要是指为保证输血安全、预防交叉感染,从而对血型、凝血、感染等进行检测。其常规检测指标包括血型、交叉配血、纤维蛋白原、病毒性肝炎、人类免疫缺陷病毒和梅毒等。传统的临床输血前检测技术以免疫分析为主。随着临床救治需求由院内向现场救治拓展,电化学传感技术、微流控技术、光谱技术等新技术也逐步发展用于输血前快速检测。基于此,本文综述了不同输血前检测技术的应用场景和优缺点,分析了系列新技术在输血前检测中的应用及未来发展趋势,为推动输血前检测乃至疾病标志物快速检测技术的发展提供参考。     

微流控芯片技术在食品领域中的应用

微流控芯片技术是一种全新的微量分析技术。介绍了微流控芯片技术的基本原理、特点及分类,并深入讨论了该技术在食品安全、营养、加工和风味等食品领域中的应用,包括有害化学物质、食品添加剂、转基因食品和食源性致病微生物等的检测,营养物质和功能成分的分析鉴定,食品工艺参数的调控以及食品风味成分的检测,展望了微流控芯片技术在食品领域的广阔应用前景。     

循环肿瘤细胞捕获与检测的纳米分析技术进展

随着纳米科学技术的发展,结构可控、表面多功能化、生物相容性良好的纳米材料在生物医药领域的各个方面都具有广泛的应用.作为一种重要的血液生物学标志物,循环肿瘤细胞(CTC)是肿瘤转移的"种子",活力较强的肿瘤细胞随着血液的流动可穿出血管在远端聚集形成微小的癌栓,对CTC的检测可用于癌症的早期诊断和转移的评估.新型纳米材料以及纳米表征测量技术的应用对CTC分析技术的进步产生了巨大的影响.近年来,基于纳米材料和微流控技术对CTC的捕获和检测已成为液体活检的研究热点,这一技术也被逐步推广到临床应用中.本文对纳米材料与纳米技术在CTC的捕获和检测中所发挥的作用进行了综述,并展望了该领域生物分析的应用前景.     

电化学免疫传感器在传染病快速检测中的应用

传染病的快速检测是传染病预防控制的重要环节,其中现场快速检测对于及时有效控制传染病疫情尤为关键。相比于传统检测方法,电化学免疫传感器具有操作简单、快速、灵敏、准确、设备可小型化等优势,可用于传染病快速检测。简要综述了近年来电化学免疫传感器在传染病快速检测中的应用研究进展,重点阐述了该类传感器在现场检测中的主要贡献和不足之处,以及免疫磁分离技术与电化学传感检测相结合在传染病快速检测方面的优势。     

基于微流控芯片的细胞-细菌相互作用光电检测技术

细胞/细菌及其相互作用研究对于生命科学、药物研发、医学诊疗等领域的研究具有重要意义。微流控芯片分析技术因微环境可控、生物相容性好、检测并行性、微型化等特性，正发展成为细胞/细菌及其相互作用研究的高效手段。本文在简要介绍基于微流控芯片分析技术的细胞-细菌分析方法和技术基础之上，对微流控芯片上细胞-细菌相互作用模型的建立进行了讨论，重点针对细胞-细菌及其相互作用过程的芯片检测进行了综述，尤其对芯片集成的光电检测技术及其测试效果进行总结和比较。通过芯片集成微流体控制、多种光电传感监测模块，使微流控芯片分析技术成为细胞/细菌及其相互作用过程分析和检测的支撑平台和优势手段。最后，对微流控光电检测技术在细胞-细菌相互作用检测中面临的挑战及发展趋势进行了讨论和展望。     

构建基于磁珠微流控芯片的iFlora遗传信息高效获取系统

磁珠以其比表面积大、易与生物分子耦联、操控方便等优点,在生命科学中得到了广泛应用。随着微机电系统（MicroElectroMechanicalSystems,MEMS）技术的发展,将磁珠应用到微流控芯片中构建磁珠微流控分析系统,为生物样品分离、检测提供了一种全新方法。新一代植物志iFlora融入现代DNA测序技术．应用高速发展的信息、网络技术及云计算分析平台,收集、整合和管理植物物种相关信息,以实现物种智能鉴定和数据提取,而包括DNA条形码在内的遗传信息及其获取技术在iFlora中的作用至关重要。本文重点概述了基于纳米磁珠的微流控芯片技术及其在分子生物学领域中的应用,提出构建基于纳米磁珠微流控芯片的iFlora遗传信息采集系统,在微芯片上完成从DNA提取到测序全过程,实现物种遗传信息的快速、高效获取。     

基于微流控芯片的秀丽隐杆线虫的研究进展

微流控芯片技术作为近年来最前沿的分析技术之一,已经在化学、生物学、医药学等研究领域取得了突破性的进展.微流控芯片具有高通量、微型化和多功能集成化等独特优势,已经成为生物医学研究的新平台之一,被越来越多地应用于秀丽隐杆线虫的研究.综述了基于微流控芯片上的秀丽隐杆线虫在生物医学领域中的研究进展,侧重介绍了微流控芯片在线虫的自动化固定、行为学、衰老与发育学、神经学、药物筛选及基因筛选等六大方面所取得的最新进展,并展望了微流控芯片的应用前景.     

Rapid analysis of fatty acid-binding proteins with immunosensors and immunotests for early monitoring of tissue injury

Fatty acid-binding protein (FABP) holds promise for early detection of tissue injury. This small protein (15 kD) appears earlier in the blood than large proteins after cell damage. Combined its characteristics of high concentration tissue contents and low normal plasma values provide the possibility of a rapid rise above the respective reference values, and thus an early indication of the appearance of tissue injury. A general review was presented on the current status of different types of FABP for the detection of tissue injury in patients with myocardial injury, brain injury and also in athletes or horses with skeletal muscle injury.

To take full advantage of the characteristics of the early marker FABP, rapid analysis is a crucial parameter. In this review, an overview of the development of immunoassay for the quantification of FABP in buffer, plasma or whole blood was outlined. The characteristics of different FABP immunosensors and immunotests were described. The feasibility of these immunoassays to be used in routine clinical practice and in emergency case was also discussed.

Nowadays, the improved automated immunoassays (e.g. a microparticle-enhanced turbidimetric immunoassay), less time-consuming bedside immunosensors and immunotests (e.g. a one-step FABP lateral flow immunotest), are the main advance technology in point-of-care testing. With these point-of-care tests, the application of FABP as an early tissue injury marker has a great potential for many clinical purposes.     

Fish-on-a-chip: a sensitive detection microfluidic system for alzheimer's disease

Microfluidics has become an important tool in diagnosing many diseases, including neurological and genetic disorders. Alzheimer's disease (AD) is a neurodegenerative disease that irreversibly and progressively destroys memory, language ability, and thinking skills. Commonly, detection of AD is expensive and complex. Fluorescence in situ hybridization (FISH)-based microfluidic chip platform is capable of diagnosing AD at an early stage and they are effective tools for the diagnosis with low cost, high speed, and high sensitivity. In this review, we tried to provide basic information on the diagnosis of AD via FISH-based microfluidics. Different sample preparations using a microfluidic chip for diagnosis of AD are highlighted. Moreover, rapid innovations in nanotechnology for diagnosis are explained. This review will provide information on dynamic quantification methods for the diagnosis and treatment of AD. The knowledge provided in this review will help develop new integration diagnostic techniques based on FISH and microfluidics.     

肿瘤蛋白标志物分析技术研究进展

目前,我国的肿瘤发病率为285.91/10万,平均每分钟有6人被诊断为恶性肿瘤,因此肿瘤的早期诊断的重要性显而易见。随着分子生物学、免疫学诊断技术的飞速发展,寻找肿瘤诊断和预后的特异性标志物已成为近期研究的热点。蛋白质是生命活动中多项功能的执行者,可以直接反映基因给予的信息。相比于基因,蛋白质的表达谱更可以直接地反映功能机制。针对蛋白质的生物特性,本文总结了目前临床中筛查肿瘤蛋白标志物分析技术,如双向凝胶电泳、基质辅助激光解吸/电离-飞行时间质谱、表面增强激光解吸/电离-飞行时间质谱、蛋白质芯片技术,并对未来肿瘤蛋白标志物筛查提出新的展望。     

Ultra-sensitive immunosensor for beta-amyloid (1-42) using scanning tunneling microscopy-based electrical detection

An ultra-sensitive immunosensor for beta-amyloid is crucial because beta-amyloid is an important challenging marker to detect for early diagnosis of Alzheimer's disease. In this study, a vertically configured electrical detection system was developed based on scanning tunneling microscopy (STM) to detect antigen-antibody binding events. This technique could be used to easily construct a multiple measurement system in a biochip. We utilized immunocomplexes comprised of the model protein, beta-amyloid (1-42), corresponding antibody fragments, and gold (Au) nanoparticles-antibody conjugates for an immunosensor for Alzheimer's disease. The electrical tunneling current between the STM tip and these complexes exhibited a peak-like pulse, the frequency of which depended on the density of the bound complexes on the surface. We could therefore quantitatively measure beta-amyloid (1-42) concentrations as low as 10fg/mL using periodogram analysis of the peak frequency. Since this method accurately quantified much smaller amounts of beta-amyloid (1-42) than traditional immunosensors, this system shows promise as an ultra-sensitive immunodetection method.     

阿尔茨海默病体外诊断纳米技术

阿尔茨海默病(Alzheimer's disease,AD)是一种最常见的神经退行性疾病.AD的精准诊断技术,特别是早期诊断技术是临床亟需的.近年来,以生物标志物为基础的非侵入性体外诊断技术发展迅速,特别是利用纳米材料和纳米技术的高表面活性、独特的光电特性、生物相容性好、易于表面修饰、小型化、集成化等特点,发展了新型的...     

A novel microfluidic immunoassay system based on electrochemical immunosensors: an application for the detection of NT-proBNP in whole blood

Electrochemical immunosensors have attracted great interest in the search for a selective, simple and reliable system for molecular recognition. Presently, electrochemical immunosensors have been widely studied for biomedical molecular's detection, but the regeneration of these immunosensors has restricted their wide application. To prepare a regeneration-free immunosensor, which may be more suitable for clinical determination, a repeatable immunoassay system was developed based on an electrochemical immunosensor with magnetic nanoparticles, biotin-avidin system (BAS) and Fab antibodies for the heart failure markers aminoterminal pro-brain natriuretic peptides (NT-proBNP). At the same time, a microfluidic system was combined into the proposed system, which enabled continuous determination. Using NT-proBNP as a model system, the proposed immunosensor exhibited rapid and sensitive amperometric response to NT-proBNP with good selectivity, stability, and a wide linear range (0.005-1.67 ng/mL and 1.67-4 ng/mL with a detection limit of 0.003 ng/mL under optimal conditions). Importantly, the proposed immunosensor was also suitable for the detection of other proteins and provided new opportunities for disease diagnosis.     

Electrochemical and chemiluminescent immunosensors for tumor markers

The determination of serum tumor markers plays an important role in clinical diagnoses for the patients with certain tumor-associated disease. Although many commercial kits have been applied in clinical immunoassays, conventional methods always have some disadvantages, resulting in the need of other new, efficient, and easily automated methods. Immunosensors, considered as a major development in immunochemical field, have attracted considerable attention. With the aim of rapid screening, many immunosensors that are small, semi-automated and portable are being developed. This brief review focuses on the current research of immunosensors for tumor markers based on the electrochemical and chemiluminescent detection with emphasis on recent advances, challenges, and trends. The works on series of novel immunosensors developed for the determination of tumor markers in our group in the last few years are also introduced.     

Development of urinary albumin immunosensor based on colloidal AuNP and PVA

The development of immunosensors with high sensitivity and specificity in detecting the pathogenic or physiologically relevant molecules in the body, offers a powerful opportunity in early diagnosis and treatment of diseases. In this study, we developed a new competitive immunosensor with employing antibody (Ab) labeled AuNP (Ab-AuNP) and PVA modified screen-printed carbon electrode (SPCE) surface to detect the urine albumin. Field emission scanning electron microscopy (FE-SEM) of modified electrode showed a suitable and stable attachment between HSA antigen- mAb and AuNP. Cyclic voltammetric (CV) method demonstrated that modification process was well performed. Electrochemical measurements including differential pulse voltammetry (DPV) and square wave voltammetry (SWV) were employed for quantitative antigen detection. The electrochemical measurements performed with other proteins mixed with samples demonstrated a high specificity and selectivity for this biosensor in detecting the HSA. In optimal conditions, the immunosensor could detect HSA in a high linear range (from 2.5 to 200 μg/mL) with a low detection limit of 25 ng/mL. This new strategy could be improved and applied to detect the other antigen.