光学分析方法在DNA检测中的应用

介绍了用于DNA检测的各种光学分析方法及其原理,主要包括荧光法、化学发光法、光纤传感法、比色法、表面等离子共振法以及其他光学衍生方法。     

单碱基多样性的非测序检测

单碱基多样性（SNP）是最常见的基因突变形式之一,经研究证明与很多疾病相关。虽然测序是检测SNP的重要方法,但其需要检测仪器,且检测时间较长,限制了其临床应用。本文综述了SNP的常见非测序分析方法。首先讨论了检测的热力学问题,并归纳了主要的检测策略：基于杂交的检测,基于链取代反应的检测和酶介导的检测。在三维均相检测方法中,主要介绍了不同信号开关策略,如荧光开关、酶识别开关和场效应开关。三维原位检测不仅能检测SNP,还能提供其细胞定位信息,在细胞异质性较高时更具优势。二维界面检测的识别反应速率和杂交效率受到一定影响,但界面检测能进一步减小干扰,亦便于实现高通量检测。以DNA正四面体探针界面为代表的改良界面具有优良的灵敏度和特异性。同时本文亦讨论了现有方法的局限性,并对SNP非测序检测研究进行展望。     

Study of the peak broadening due to detection in the electrophoretic separation of DNA by CE and microchip CE and the application of image sensor for ultra‐small detection cell length

Narrow peaks are important to high‐resolution and high‐speed separation of DNA fragments by capillary electrophoresis and microchip capillary electrophoresis. Detection cell length is one of the broadening factors, which is often ignored in experiments. However, is it always safe to neglect detection cell length under any condition? To answer this question, we investigated the influence of detection cell length by simulation and experiments. A parameter named as detection cell length ratio was proposed to directly compare the detection cell length and the spatial length of sample band. Electrophoretic peaks generated by various detection cell length ratios were analyzed. A simple rule to evaluate the peak broadening due to detection cell length was obtained. The current states of the detection cell length of detection system and their reliabilities in capillary electrophoresis and microchip capillary electrophoresis were analyzed. Microchip capillary electrophoresis detection with an ultra‐small detection cell length of 0.36 μm was easily achieved by using an image sensor.     

毛细管电泳与芯片毛细管电泳的双检测技术

综述了毛细管电泳和芯片毛细管电泳的3种双检测技术,包括荧光-散射等光学双检测技术、安培-非接触电导等电化学双检测技术和荧光-非接触电导等光电联用双检测技术.介绍了3种双检测方法的仪器的检测原理及应用,并展望了双检测技术的发展前景.引用文献54篇.     

Fluorescence-based nucleic acid detection and microarrays

Recent analytical innovations for nucleic acid detection have revolutionized the biological sciences. Single nucleic acid sequence detection methods have been expanded to incorporate multiplexed detection strategies. A variety of nucleic acid detection formats are now available that can address high throughput genomic interrogation. Many of these parallel detection platforms or arrays, employ fluorescence as the signaling method. Fluorescence-based assays offer many advantages, including increased sensitivity, safety and multiplexing capabilities, as well as the ability to measure multiple fluorescence properties. Multiplexed microarray platforms provide parallel detection capabilities capable of measuring thousands of simultaneous responses. This review will discuss both single target detection and microarray applications with a focus on gene expression and pathogenic microorganism (PM) detection.     

Comparison of pulsed amperometric detection and integrated voltammetric detection for inorganic sulfur compounds in liquid chromatography

A Variety of potential–time waveforms are useful in pulsed electrochemical detection (PED) when applied for the amperometric detection of numerous polar organic compounds following their separation by liquid chromatography (LC). Here, we compare the waveforms for pulsed amperometric detection (PAD) and integrated voltammetric detection (IVD) applied for detection of organosulfur compounds at Au electrodes in acidic media. In PAD waveforms, electrodes response is measured at a constant detection potentials. In IVD waveforms, electrodes current is integrated throughout a fast cyclic scan of the detection potential. As a consequence of this difference in detection strategy, the background signal for IVD is significantly smaller for PAD in the detection of organosulfur compounds whose response mechanisms require the concomitant formation of surface oxides on Au electrodes. Furthermore, in comparison to Pad, IVD has a larger sensitivity and a diminished system peak from 0₂ dissolved in the sample. Use of a preadsorption step increases detection sensitivity in both PAD and IVD. The limit of detection (S/N=3)for cysteine in LC-IVD is ca. 6 nM for a 50-μl injection (i.e., 300 fmol) using a detection waveform that includes a 1000-ms preadsorption period.     

基于拉曼光谱的细菌检测研究进展

细菌是一种与人类生命活动息息相关的微生物, 其快速、 高灵敏检测对重大传染性疾病的防控至关重要. 本文介绍了拉曼光谱用于细菌检测的基本原理, 综述了3种拉曼光谱用于细菌检测的主要方式, 包括细菌组成成分检测、 细菌代谢物检测以及基于拉曼探针标记的检测模式, 并对各种拉曼检测方法进行了分析比较. 最后, 展望了拉曼光谱在细菌检测领域的发展前景, 并提出了5条建议.     

离子色谱中的安培检测方法及其应用

介绍了离子色谱中的安培检测方法(包括恒电位安培检测法、脉冲安培检测法和积分脉冲安培检测法)的原理和应用。脉冲安培检测法与高效阴离子交换色谱结合(HPAEC-PAD)是一种新的分析糖类化合物的方法;积分脉冲安培检测法与高效阴离子交换色谱结合(HPAEC-IPAD)是一种新的氨基酸分析方法。     

Separation of perfluorocarboxylic acids using capillary electrophoresis with UV detection

A capillary electrophoretic method with UV detection for separation and quantitation of perfluorocarboxylic acids (PFCAs) from C6-PFCA to C12-PFCA has been developed. The optimization of measurement conditions included the choice of the most appropriate type and concentration of buffer in the background electrolyte (BGE), as well as the type and the content of an organic modifier. The optimal separation of investigated PFCAs was achieved with 50 mM phosphate buffer and 40% isopropanol in the BGE using direct UV detection. The optimum wavelength for direct UV detection was optimized at 190 nm. For indirect detection, several chromophores were studied. Five mM 3,5-Dinitrobenzoic acid (3,5-DNBA) in 20 mM phosphate buffer BGE and indirect UV detection at 280 nm gave the optimal detection and separation performance for the investigated PFCAs. The possibility of on-line preconcentration of solutes by stacking has been examined for indirect detection. The detection limits (LODs) determined for direct UV detection ranged from 2 microg/mL for C6-PFCA to 33 microg/mL for C12-PFCA. The LODs obtained for indirect UV detection were comparable to those obtained for direct UV detection.     

Label-free fluorescence detection in capillary and microchip electrophoresis

Herein, we summarize the current status of native fluorescence detection in microchannel electrophoresis, with a strong focus on chip-based systems. Fluorescence detection is a powerful technique with unsurpassed sensitivity down to the single-molecule level. Accordingly fluorescence detection is attractive in combination with miniaturised separation techniques. A drawback is, however, the need to derivatize most analytes prior to analysis. This can often be circumvented by utilising excitation light in the UV spectral range in order to excite intrinsic fluorescence. As sensitive absorbance detection is challenging in chip-based systems, deep-UV fluorescence detection is currently one of the most general optical detection techniques in microchip electrophoresis, which is especially attractive for the detection of unlabelled proteins. This review gives an overview of research on native fluorescence detection in capillary (CE) and microchip electrophoresis (MCE) between 1998 and 2008. It discusses material aspects of native fluorescence detection and the instrumentation used, with particular focus on the detector design. Newer developments, featured techniques, and their prospects in the future are also included. In the last section, applications in bioanalysis, drug determination, and environmental analysis are reviewed with regard to limits of detection.     

Element-selective atomic emission detection for capillary GC of metal-containing compounds

Element-selective GC detection by microwave-induced plasma atomic emission spectroscopy has been used to examine a wide variety of compounds containing metals, non-metals, and metalloids. “Recipes”, or new selective detection schemes for use with the software of the computer-controlled system, have been developed for the selective detection of boron, aluminum, gallium, titanium, vanadium, chromium, manganese, rhenium, palladium, and platinum. Figures of merit including limits of detection, linear dynamic range, and spectral selectivity over carbon have been established for most of these elements. Gas chromatography – atomic emission detection (GC-AED) has been applied to the selective detection of vanadium, nickel, and iron in metalloporphyrins present in crude oil, manganese-selective detection of methylcyclopentadienylmanganese tricarbonyl (MMT) in gasoline, and titanium-selective detection of reaction mixtures containing titanium catalysts or titanium boride molecular precursors.     

毛细管电泳安培型电化学检测法

本文评述了近年来安培型电化学检测法在毛细管电泳中的发展概况，归纳了二种检测方式及其装置的结构特点。阐述了金属电极、液膜电极和修饰电极等在电化学检测应用研究中的作用以及此方法在微区分析、活体分析中的重要意义，并展望了其发展前景。     

Detection of carbohydrates in capillary electrophoresis

This review focuses on recent developments in sensitive detection modes for carbohydrates after separation by capillary electrophoretic methods. To bring detection sensitivity for carbohydrates analysis in line with current methods in protein sequencing, concentration detection limits of 10⁻⁶ molar or better are requires. A discussion of mass detection limits and concentration detection limits is followed by an overview of detection modes for natural and labeled carbohydrates. Amperometric detection and UV and laser-induced fluorescence detection after reductive amination, in particular with 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS), are discussed in more detail. Finally, the paper outlines developments to be expected in the near future, focusing on the needs in glycobiology such as improved sensitivity and selectivity.     

Laser-based refractive index detection for micro-channels

Laser-based refractive index detection, based upon refractive index changes in micro-channels, is a kind of laser-based optical determination for solutions. As it is capable of label-free determination, it has been regarded as a considerable growth prospect. The key aspect in related researches lay in the development of novel detection configurations, which would possibly result in better sensitivity. Over the past decade, micro-channel laser-based refractive index detection has been significantly improved, resulting in traversed, hologram-based, back-scattered, retro-reflected interference, optical ring resonant, etc., detection configurations. Moreover, laser-based refractive index detection has been combined with other laser-based detection strategies, in order to pursue a universal and sensitive detection. And last but not least, some utilizations of laser-based refractive index detection have been reported, while both advantages and drawbacks exist. In this paper, laser-based refractive index detection for micro-channels will be reviewed in proceeding sequence.     

“Getting the best sensitivity from on-capillary fluorescence detection in capillary electrophoresis” – A tutorial

Capillary electrophoresis with Laser-Induced Fluorescence (CE-LIF) detection is being applied to new analytical problems which challenge both the power of CE separation and the sensitivity of LIF detection. On-capillary LIF detection is much more practical than post-capillary detection in a sheath-flow cell. Therefore, commercial CE instruments utilize solely on-capillary CE-LIF detection with a Limit of Detection (LOD) in the nM range, while there are multiple applications of CE-LIF that require pM or lower LODs. This tutorial analyzes all aspects of on-capillary LIF detection in CE in an attempt to identify means for improving LOD of CE-LIF with on-capillary detection. We consider principles of signal enhancement and noise reduction, as well as relevant areas of fluorophore photochemistry and fluorescent microscopy.     

High-performance liquid chromatographic separation and detection methods for anabolic compounds.

The role of high-performance liquid chromatography (HPLC) in methods of analysis for anabolic compounds in biological samples is reviewed. Special attention is given to both the separation and detection of anabolic compounds. A distinction is made between on-line detection systems, such as ultraviolet detection and diode-array detection, and off-line detection methods with special emphasis on immunochemical detection methods using non-isotopic labels. A number of applications are given to elucidate the possibilities of HPLC in the analysis of anabolic compounds.     

功能化纳米材料的制备及在食品安全检测中的应用研究进展

纳米材料研究的飞速发展为解决食品安全检测所面临的问题提供了很好的理论基础和技术支持。功能化纳米材料与传统检测方法的结合,衍生出具有高灵敏、高通量、快速检测的方法,在食品中有毒有害微生物、化学残留物以及毒素的检测方面的应用正逐渐引起人们的重视。本文对基于免疫方法的常见功能化纳米材料在食品安全检测中的应用的研究近况进行了综述。     

Capillary electrophoresis with end-capillary potentiometric detection using a copper electrode.

Potentiometric end-capillary detection in capillary electrophoresis has the advantage of relatively easy miniaturisation without having to compromise the concentration sensitivity. Potentiometric end-capillary detection using a copper electrode is also attractive because of the sensitive detection of many inorganic and organic UV-transparent ions and the ability to work in both direct and indirect mode. In this work, detection of a number of common anions in a tartrate electrolyte at pH 3 was studied. The influence of the end-capillary detection geometry on the detection performance was investigated. An end-capillary detection cell allowing the separation capillary to be changed without the need to realign the detection electrode was constructed and fitted into a commercial CE apparatus. Under the optimal configuration, which was a 25 microm diameter copper electrode aligned coaxially with a 25 microm capillary and positioned at a distance of about 25 microm from the capillary end, excellent peak shapes were achieved and comparison with simultaneous on-capillary photometric detection showed no additional peak broadening. Good sensitivity was obtained, resulting in concentration limits of detection (LODs) in the low microM range and mass LODs in the low amol range. Examples of separations of inorganic and organic anions are presented and the analytical potential of the detection method is assessed.     

纳米材料应用于DNA检测领域的研究进展

本文主要评述了近年来纳米材料在除了PCR领域以外的DNA检测方面的研究进展.对以纳米材料(纳米粒子、纳米纤维、纳米线、纳米管)为单元,或以纳米器件的制备为实验方法而开展的DNA检测方面的工作进行了介绍.研究表明,基于纳米材料的DNA检测法,无论是在定位、可视化还是多重检测等方面都比传统PCR技术的检测方法表现出其自身的...     

An evaluation of the experimental approaches to detection of small ions in CE

This review points out some important trends in the development of the detection techniques for small ions in CE. On the basis of selected literature references it briefly discusses some general requirements on detection techniques in CE. Various optical measurements, mass spectrometric approaches and electrochemical detection techniques are dealt with. Some specific features of microchip CE separation and detection are pointed out and possibilities of dual detection are mentioned. The principal parameters of the above detection techniques are then briefly compared.