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1.
刘巨  梁涛波  许恒毅 《分析测试学报》2020,39(12):1556-1560
滚环扩增(RCA)是一种等温核酸扩增技术,因其具有扩增效率高、保真度高等特点,在生物传感器领域得到了广泛的应用。该文介绍了RCA技术的基本原理、RCA环状模板环化方式和RCA的扩增类型,并对基于RCA技术的荧光、比色以及电化学生物传感器进行了介绍,旨在为RCA技术在生物传感器领域的进一步发展和应用提供参考。  相似文献   

2.
DNA电化学生物传感器是一类以DNA为敏感元件或检测对象,将核酸分子特异性识别过程中产生的信号通过换能器转化为电信号,从而实现对目标物定性或定量检测的传感器,具有响应速度快、操作简单、选择性好、灵敏度高、检测成本低等优点,实现了多领域中重金属、真菌毒素、核酸等的快速实时检测。介绍了DNA电化学生物传感器的组装单元、电化学指示剂类型,以DNA二级构型角度综述了DNA电化学生物传感器的四大类特殊结构,并汇总其在临床、中医药、生态环境保护及食品安全等领域中重金属的检测应用研究,对新型DNA电化学生物传感器的设计与其在更多领域的拓展应用提供借鉴价值。  相似文献   

3.
基于核酸适体的电化学生物传感器*   总被引:3,自引:0,他引:3  
核酸适体是一类体外筛选的、可与目标分子高效、高特异亲合的RNA或DNA寡核苷酸片段,与常规识别分子(如抗体等)相比,核酸适体作为一类新型识别分子具有明显特色和优势,已被广泛应用于生物传感等分子识别和应用研究领域。本文就基于核酸适体的电化学生物传感器(标记型和非标记型)的近期进展作简要评述,包括适体简介、标记型(“信号衰减”型、“信号增强”型、酶标记型和纳米粒子标记型)和非标记型电化学适体生物传感器等内容。  相似文献   

4.
核酸适配体是指能与特定靶分子结合的寡核苷酸链。因其具有易修饰、易合成和低免疫原性等特点,作为生物传感器的靶向元件,已广泛应用于各种生物标志物检测技术中。本文综述了近年来基于核酸适配体的生物标志物的检测技术在癌症、心血管疾病、阿尔兹海默症、抑郁症等多个疾病领域的诊断研究进展,列举核酸适配体新兴技术的创新应用,以期为生物标志物的检测提供新思路,也有望为相关疾病的早期诊断和治疗提供参考。  相似文献   

5.
信号放大是新型生物传感分析过程中重要的环节。核酸介导的信号放大技术凭借核酸材料灵活的结构设计、低成本和易于制备等特点,在生物传感快速检测技术的开发上逐渐发展成为一项重要的分支,广泛应用于食品、环境和医药等新型检测方法开发。介绍了传统和新型核酸扩增技术、生物条形码和DNA walker等信号放大机理和应用,同时进一步综述核酸信号放大技术结合光学生物传感在食品污染物中检测的应用,如化学污染物、毒素类污染物、和重金属污染物等,并对核酸介导的信号放大技术在食品污染物检测中的问题和前景进行讨论。  相似文献   

6.
张晗  丁家旺  秦伟 《化学进展》2021,33(10):1756-1765
多肽具有分子量小、易于合成、生物兼容性好、稳定性高及序列灵活多样等优点。因此,多肽作为新型生物识别元件,已被广泛应用于生物传感器的构建。电化学分析灵敏度高、准确度好、设备简单、检测范围广且易于操作。本文介绍了基于多肽识别的电化学生物传感器技术,包括多肽的修饰与固定化、多肽与待测物的识别及检测原理;综述了近五年多肽电化学生物传感器对重金属离子、小分子、蛋白质、细菌和病毒的检测;展望了肽基电化学生物传感器的发展趋势。  相似文献   

7.
近年来,基于离子通道的电化学检测技术备受关注,目前该技术已广泛应用于DNA测序、分子间相互作用测定以及无机离子、生物分子等检测中。本文从电化学检测方法的角度,综述了近年来基于离子通道的电流型、阻抗型以及电位型检测技术在化学与生物传感中的应用研究进展,重点介绍了此技术的作用原理及传感器构建方法,并对离子通道基电化学传感技术进行了展望。  相似文献   

8.
生物电化学简介   总被引:4,自引:0,他引:4  
简单介绍了生物电化学研究领域的概况。包括:生物膜与生物界面模拟研究(SAM膜模拟生物膜的电化学、液/液界面模拟生物膜的电化学),用于生命科学的电化学技术(电脉冲基因直接导入、电场加速作物生长、癌症的电化学疗法、电化学控制药物释放、在体研究的电化学方法、生物分子的电化学行为)和电化学生物传感器(酶电极传感器、微生物电极传感器、电化学免疫传感器、组织电极与细胞器电极传感器、电化学DNA传感器)。  相似文献   

9.
富鸟嘌呤的单链核酸在一定条件下可通过折叠形成特殊的G-四聚体和G-三聚体核酸高级结构。该结构和功能具有丰富的多样性,可作为多功能的信号元件在各类生物传感器的构建中发挥作用。电化学生物传感器因其具有灵敏度高、易于微型化、成本低廉及分析速度快等优点,被广泛应用于各类目标物的分析检测中。近年来,随着现场检测、快速检测成为体外诊断的一大热点,越来越多的研究人员聚焦于开发G-四聚体和G-三聚体在电化学生物传感器中的应用。本文针对G-四聚体和G-三聚体的电化学生物传感器在金属离子、有机小分子、核酸和蛋白大分子及酶活性等的最新分析检测方法的研究进行了综述。  相似文献   

10.
生物电分析化学是一门利用生物分子作为识别元件、通过生物反应后电极过程产生的信号的变化对未知物质进行定性、定量分析的学科.目前常用的生物识别元件有酶、核酸(包括核酸适配体)、抗体、受体等,而可供检测的信号广义上包括电流、电阻、光电流和电化学发光等.在过去5年的工作中,我们针对生命科学、临床检验和环境监测等实际应用领域,分别研究了电化学酶传感器、光电化学核酸损伤传感器和电化学发光免疫检测的原理与技术,并研制了相应的检测仪器.  相似文献   

11.
The most popular in vitro nucleic acid amplification techniques like polymerase chain reaction (PCR) including real-time PCR are costly and require thermocycling, rendering them unsuitable for uses at point-of-care. Highly efficient in vitro nucleic acid amplification techniques using simple, portable and low-cost instruments are crucial in disease diagnosis, mutation detection and biodefense. Toward this goal, isothermal amplification techniques that represent a group of attractive in vitro nucleic acid amplification techniques for bioanalysis have been developed. Unlike PCR where polymerases are easily deactivated by thermally labile constituents in a sample, some of the isothermal nucleic acid amplification techniques, such as helicase-dependent amplification and nucleic acid sequence-based amplification, enable the detection of bioanalytes with much simplified protocols and with minimal sample preparations since the entire amplification processes are performed isothermally. This review focuses on the isothermal nucleic acid amplification techniques and their applications in bioanalytical chemistry. Starting off from their amplification mechanisms and significant properties, the adoption of isothermal amplification techniques in bioanalytical chemistry and their future perspectives are discussed. Representative examples illustrating the performance and advantages of each isothermal amplification technique are discussed along with some discussion on the advantages and disadvantages of each technique.  相似文献   

12.
X Fang  H Zhang  F Zhang  F Jing  H Mao  Q Jin  J Zhao 《Lab on a chip》2012,12(17):3190-3196
This paper reports the design and implementation of a contactless conductivity detection system which combines a thermal control cell, a data processing system and an electrochemical (EC) cell for label-free isothermal nucleic acid amplification and real-time monitoring. The EC cell consists of a microchamber and interdigitated electrodes as the contactless conductivity biosensor with a cover slip as insulation. In our work, contactless EC measurements, the effects of trehalose on amplification, and chip surface treatment are investigated. With the superior performance of the biosensor, the device can detect the amount of pure DNA at concentrations less than 0.1 pg μl(-1). The EC cell, integrated with a heater and a temperature sensor, has successfully implemented nicking-based strand-displacement amplification at an initial concentration of 2.5 μM and the yields are monitored directly (dismissing the use of probes or labels) on-line. This contactless detector carries important advantages: high anti-interference capability, long detector life, high reusability and low cost. In addition, the small size, low power consumption and portability of the detection cell give the system the potential to be highly integrated for use in field service and point of care applications.  相似文献   

13.
Kivlehan F  Mavré F  Talini L  Limoges B  Marchal D 《The Analyst》2011,136(18):3635-3642
We described an electrochemical method to monitor in real-time the isothermal helicase-dependent amplification of nucleic acids. The principle of detection is simple and well-adapted to the development of portable, easy-to-use and inexpensive nucleic acids detection technologies. It consists of monitoring a decrease in the electrochemical current response of a reporter DNA intercalating redox probe during the isothermal DNA amplification. The method offers the possibility to quantitatively analyze target nucleic acids in less than one hour at a single constant temperature, and to perform at the end of the isothermal amplification a DNA melt curve analysis for differentiating between specific and non-specific amplifications. To illustrate the potentialities of this approach for the development of a simple, robust and low-cost instrument with high throughput capability, the method was validated with an electrochemical system capable of monitoring up to 48 real-time isothermal HDA reactions simultaneously in a disposable microplate consisting of 48-electrochemical microwells. Results obtained with this approach are comparable to that obtained with a well-established but more sophisticated and expensive fluorescence-based method. This makes for a promising alternative detection method not only for real-time isothermal helicase-dependent amplification of nucleic acid, but also for other isothermal DNA amplification strategies.  相似文献   

14.
A multi-analyte biosensor based on nucleic acid hybridization and liposome signal amplification was developed for the rapid serotype-specific detection of Dengue virus. After RNA amplification, detection of Dengue virus specific serotypes can be accomplished using a single analysis within 25 min. The multi-analyte biosensor is based on single-analyte assays (see Baeumner et al (2002) Anal Chem 74:1442–1448) developed earlier in which four analyses were required for specific serotype identification of Dengue virus samples. The multi-analyte biosensor employs generic and serotype-specific DNA probes, which hybridize with Dengue RNA that is amplified by the isothermal nucleic acid sequence based amplification (NASBA) reaction. The generic probe (reporter probe) is coupled to dye-entrapping liposomes and can hybridize to all four Dengue serotypes, while the serotype-specific probes (capture probes) are immobilized through biotin–streptavidin interaction on the surface of a polyethersulfone membrane strip in separate locations. A mixture of amplified Dengue virus RNA sequences and liposomes is applied to the membrane and allowed to migrate up along the test strip. After the liposome-target sequence complexes hybridize to the specific probes immobilized in the capture zones of the membrane strip, the Dengue serotype present in the sample can be determined. The amount of liposomes immobilized in the various capture zones directly correlates to the amount of viral RNA in the sample and can be quantified by a portable reflectometer. The specific arrangement of the capture zones and the use of unlabeled oligonucleotides (cold probes) enabled us to dramatically reduce the cross-reactivity of Dengue virus serotypes. Therefore, a single biosensor can be used to detect the exact Dengue serotype present in the sample. In addition, the biosensor can simultaneously detect two serotypes and so it is useful for the identification of possible concurrent infections found in clinical samples. The various biosensor components have been optimized with respect to specificity and sensitivity, and the system has been ultimately tested using blind coded samples. The biosensor demonstrated 92% reliability in Dengue serotype determination. Following isothermal amplification of the target sequences, the biosensor had a detection limit of 50 RNA molecules for serotype 2, 500 RNA molecules for serotypes 3 and 4, and 50,000 molecules for serotype 1. The multi-analyte biosensor is portable, inexpensive, and very easy to use and represents an alternative to current detection methods coupled with nucleic acid amplification reactions such as electrochemiluminescence, or those based on more expensive and time consuming methods such as ELISA or tissue culture.  相似文献   

15.
Miniaturized isothermal nucleic acid amplification, a review   总被引:1,自引:0,他引:1  
Asiello PJ  Baeumner AJ 《Lab on a chip》2011,11(8):1420-1430
Micro-Total Analysis Systems (μTAS) for use in on-site rapid detection of DNA or RNA are increasingly being developed. Here, amplification of the target sequence is key to increasing sensitivity, enabling single-cell and few-copy nucleic acid detection. The several advantages to miniaturizing amplification reactions and coupling them with sample preparation and detection on the same chip are well known and include fewer manual steps, preventing contamination, and significantly reducing the volume of expensive reagents. To-date, the majority of miniaturized systems for nucleic acid analysis have used the polymerase chain reaction (PCR) for amplification and those systems are covered in previous reviews. This review provides a thorough overview of miniaturized analysis systems using alternatives to PCR, specifically isothermal amplification reactions. With no need for thermal cycling, isothermal microsystems can be designed to be simple and low-energy consuming and therefore may outperform PCR in portable, battery-operated detection systems in the future. The main isothermal methods as miniaturized systems reviewed here include nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP), helicase-dependent amplification (HDA), rolling circle amplification (RCA), and strand displacement amplification (SDA). Also, important design criteria for the miniaturized devices are discussed. Finally, the potential of miniaturization of some new isothermal methods such as the exponential amplification reaction (EXPAR), isothermal and chimeric primer-initiated amplification of nucleic acids (ICANs), signal-mediated amplification of RNA technology (SMART) and others is presented.  相似文献   

16.
林雪霞  王晨境  林金明 《色谱》2020,38(10):1179-1188
人乳头瘤病毒(human papillomavirus,HPV)是一种常见的球形DNA病毒,目前已报道其可以导致6种类型的癌症发生,因此HPV病毒检测方法的研究引起了人们的重视。芯片毛细管电泳(MCE),作为一种芯片实验设备,结合各种信号放大技术为HPV分型检测提供了简单、快速、高灵敏度和易便携化的检测方法。该文综述了MCE在常规HPV分型检测中的最新研究进展,主要分为MCE技术和MCE结合核酸扩增技术两个部分。综述的第一部分介绍了MCE系统、MCE芯片结构设计和电泳分离方法。典型的MCE系统包含了高压电源、分离芯片、电解液池、进样系统、检测系统等。该文还介绍了近年来应用最广泛的4种芯片通道,包括分离直通道、T型通道、蛇形通道以及双通道,并分别对它们的优缺点进行了比较。第二部分主要介绍芯片电泳在HPV检测中的应用和发展。由于MCE技术的应用,HPV目标物的分离时间,从以前的几个小时缩短到几分钟,极大地提高了分离速度。重点介绍了各种核酸扩增技术结合MCE检测HPV的方法。对聚合酶链式反应(PCR)和MCE结合用于HPV的检测技术、环介导等温扩增(LAMP)技术的HPV检测方法、基于PCR结合限制性片段长度多态性(RFLP)技术用于HPV分型的DNA检测、基于核酸序列扩增(NASBA)技术检测HPV mRNA、巢式PCR等进行了比较分析。其次,对HPV其他检测方法进行了总结,其中包括PCR结合傅里叶变换红外光谱法(FT-IR)、纳米技术、DNA探针结合电化学方法、亚铜粒子氧化还原锌掺杂的二硫化钼量子点结合T7外切酶电化学发光法和基于CRISPR/Cas12a的环介导等温扩增法。在这些非MCE方法中,电化学传感法,如阻抗法、脉冲伏安法和流动生物传感器,由于背景信号低、时间控制能力强,是一种比较理想的方法。最后,虽然近年来MCE技术得到了发展,所开发的设备得到了应用,但目前在MCE技术、方法和应用方面仍然存在一些挑战。MCE技术在HPV分型检测应用中面临的第一个挑战是,MCE本身无法对HPV核酸进行信号放大,从而不能在HPV的高灵敏和高选择性分析中得到很好的应用。第二个挑战是,虽然有一些研究者已经成功地将PCR和MCE集成在一个芯片上,但该技术的广泛应用仍面临困难,目前仍然没有真正集成的PCR-MCE芯片用于HPV检测。第三个挑战是目前MCE技术无法实现小型化、自动化器件的制造。最后,文章就MCE在HPV分型检测中开发更自动化、更快速以及更稳定可靠的检测技术提出了一些观点和见解,希望能对感兴趣的读者提供一些启发。  相似文献   

17.
In this work, we report the first electrochemistry-based real-time polymerase chain reaction technique for sequence-specific nucleic acid detection. This new technique builds upon the advantages of the well-established fluorescence-based counterpart, such as short assay time (simultaneous target DNA amplification and detection). In addition, this electrochemical approach could employ simple and miniaturizable instrumentation compared to the bulky and expensive optics required in the fluorescence-based schemes. We have demonstrated a proof-of-concept experiment showing that the utilization of solid-phase extension of the electrode surface-immobilized capture probe with Fc-dUTP during PCR resulted in the accumulation of the redox marker on the transducer surface. This new technique can be applied to a microfabricated PCR electrochemical device for point-of-care diagnostics as well as on-site environmental monitoring and biowarfare agent detection.  相似文献   

18.
Isothermal exponential amplification techniques, such as strand‐displacement amplification (SDA), rolling circle amplification (RCA), loop‐mediated isothermal amplification (LAMP), nucleic acid sequence based amplification (NASBA), helicase‐dependent amplification (HDA), and recombinase polymerase amplification (RPA), have great potential for on‐site, point‐of‐care, and in situ assay applications. These amplification techniques eliminate the need for temperature cycling, as required for the polymerase chain reaction (PCR), while achieving comparable amplification yields. We highlight here recent advances in the exponential amplification reaction (EXPAR) for the detection of nucleic acids, proteins, enzyme activities, cells, and metal ions. The incorporation of fluorescence, colorimetric, chemiluminescence, Raman, and electrochemical approaches enables the highly sensitive detection of a variety of targets. Remaining issues, such as undesirable background amplification resulting from nonspecific template interactions, must be addressed to further improve isothermal and exponential amplification techniques.  相似文献   

19.
Due to their high specificity and affinity towards various targets,along with other unique advantages such as stability and low cost,aptamers are widely applied in analytical techniques.A typical aptamerbased electrochemical biosensor is composed of a aptamer as the biological recognition element and transducer conve rting the biologic interaction into electrical signals for the quantitative measure ment of targets.Improvement of the sensitivity of a biosensor is significantly important in order to achieve the detection of biomolecules with low abundance,and different amplification strategies have been explored.The strategies either employ nanomaterials such as gold nanoparticles to construct electrodes which can trans fer the biological reactions more efficiently,or attempt to obtain enha nced signal through multi-labeled carriers or utilize enzyme mimics to catalyze redox cycling.This review discusses recent advances in signal amplification methods and their applications.Critical assessment of each method is also considered.  相似文献   

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