DNA传感器研究进展

本文概述了当前生物传感器的研究特点以及发展ＤＮＡ生物传感器的迫切性;从不同角度阐述了ＤＮＡ生物传感器的概念和研究内容;着重讨论了ＤＮＡ生物传感器的研究现状和发展趋势。文中分别对ＤＮＡ光生物传感器和ＤＮＡ压电晶体生物传感器的基本原理、特点、研究进展及存在的问题进行了分析与说明。进而,对我国ＤＮＡ生物传感器研究存在的差距和发展前景进行了简要论述。     

DNA生物传感器及其研究进展

就ＤＮＡ生物传感器的工作原理,分类、ＤＮＡ探针的固化方法,以及电化学ＤＮＡ生物传感器、光学ＤＮＡ生物传感器及压电ＤＮＡ生物传感器的研究进展、优缺点和发展趋势加以介绍。     

DNA生物传感器研究进展

本根据作用机理不同将ＤＮＡ生物传感器分为ＤＮＡ光化学传感器,ＤＮＡ电化学传感器和压电晶体传感器,并就几种方面的研究进展进行了综述。     

光纤DNA传感器

光纤ＤＮＡ传感器是ＤＮＡ生物传感器中很有潜力、极具发展前途的一种,具有重要的研究价值和诱人的应用前景。该技术灵敏度高,分析操作简便、快速,为分子生物研究提供了一种新方法,在基础医学、临床医学和预防医学各个领域具有十分广泛的应用前景。本文对光纤ＤＮＡ传感器的原理,ＤＮＡ探针在光纤表而的固定化,光纤ＤＮＡ传感器的类型等方面的研究进展作了评述。     

检测重金属离子生物传感器的研究进展北大核心CSCD

重金属污染广泛存在于环境中,它能通过生物富集作用对动植物及人类产生危害。利用分析化学方法检测重金属离子对其生物危害缺乏直接检定,生物传感器检测重金属离子吸引了越来越多的研究兴趣。对检测重金属离子的各种生物传感器进行介绍,并分析各种生物传感器的特点,指出依靠先进的生物技术研究方法和成果、了解生物传感器生物敏感材料的结构和功能,进一步开发能够自动操作的便携生物传感器是能够满足重金属污染检测要求的方法。     

基于专利分析的生物传感器发展态势研究*

生物传感器将固定化的生物敏感材料作为识别元件,具有体积小、专一性强、响应快、准确度高等特点,市场应用前景广阔。近年来,生物传感器技术发展迅速,研究成果不断涌现。为更好地了解我国生物传感器研究现状,进一步促进生物传感器行业健康快速发展,基于IncoPat专利数据库,利用ITGinsight作为文献分析工具,对全球范围内生物传感器领域的总体研究态势、区域布局、技术构成及研究热点等方面进行专利分析,并对核心专利进行识别。结果表明,我国生物传感器领域的专利数量排在世界前列,但科研实力存在区域分布不均衡、企业研发实力不强、关键核心技术不足等问题。针对这些问题,提出我国生物传感器技术的未来发展策略。     

阵列电化学生物传感器研究进展

阵列生物传感器技术作为一种高通量、快速、选择性高和集成化的分析技术,已在基因组学和蛋白质组学的研究和药物筛选、环境分析,食品分析,临床诊断等领域中得到广泛的应用.阵列生物传感器主要有阵列光学生物传感器和阵列电化学生物传感器.阵列电化学生物传感器是将生物分子识别物质如酶、抗原/抗体、DNA等固定在阵列电极上,以阵列中每根电极产生的电化学信号作为检测信号的电化学分析器件.阵列电化学生物传感器以灵敏度高、分析速度快、选择性好、易于微型化和集成化以及仪器价格低廉等特点受到了研究工作者的极大关注.本文简单介绍了阵列电化学生物传感器的原理和特点,重点评述了2005年以来阵列电化学生物传感器在单组份检测和多组份同时检测两方面的研究进展,简单讨论了阵列电化学生物传感器研究中存在的问题.     

阵列生物传感器研究进展

简要介绍了阵列生物传感器的基本原理和分类。根据换能器的不同,评述了光学、电化学、质量型、磁致阻抗等阵列生物传感器的研究进展,并对目前阵列生物传感器研究中存在的问题进行了分析。     

阵列生物传感器研究进展

简要介绍了阵列生物传感器的基本原理和分类。根据换能器的不同,评述了光学、电化学、质量型、磁致阻抗等阵列生物传感器的研究进展,并对目前阵列生物传感器研究中存在的问题进行了分析。     

分子识别诱导的蛋白质和核酸多层膜的有序组装

通过生物大分子之间的特异性结合,采用表面等离激元共振技术监测,报导了支撑于固体表面脂单层膜上进行的亲和素、生物素标记的质粒ＤＮＡ、以及从系统性红斑狼疮患者血清中获得的抗ＤＮＡ抗体多层膜的有序组装。这种生物大分子的组装技术可以用于生物传感器以检测特定的抗原抗体。     

Adaptable gene‐specific dye bias correction for two‐channel DNA microarrays

DNA microarray technology is a powerful tool for monitoring gene expression or for finding the location of DNA‐bound proteins. DNA microarrays can suffer from gene‐specific dye bias (GSDB), causing some probes to be affected more by the dye than by the sample. This results in large measurement errors, which vary considerably for different probes and also across different hybridizations. GSDB is not corrected by conventional normalization and has been difficult to address systematically because of its variance. We show that GSDB is influenced by label incorporation efficiency, explaining the variation of GSDB across different hybridizations. A correction method (Gene‐ And Slide‐Specific Correction, GASSCO) is presented, whereby sequence‐specific corrections are modulated by the overall bias of individual hybridizations. GASSCO outperforms earlier methods and works well on a variety of publically available datasets covering a range of platforms, organisms and applications, including ChIP on chip. A sequence‐based model is also presented, which predicts which probes will suffer most from GSDB, useful for microarray probe design and correction of individual hybridizations. Software implementing the method is publicly available.     

Analysis of single nucleotide incorporation reactions by capillary electrophoresis

Single nucleotide incorporation assays have been used to probe the kinetic parameters of many DNA and RNA polymerases. Traditionally, oligonucleotide primers are 5'-(32)P labeled using T4 kinase and annealed to a complementary template with a 5' overhang. To quantify the reaction kinetics, the products of the primer extension reactions are usually separated using denaturing polyacrylamide gel electrophoresis and quantified using a phosphorimager or other method to measure radioactivity. We have developed a method using a 5' fluorescently labeled oligonucleotide to examine the kinetics of single nucleotide incorporation catalyzed by recombinant human mitochondrial polymerase gamma (Pol gamma) holoenzyme. Using laser-induced fluorescence detection in the P/ACE MDQ instrument, primers 5' labeled with fluorescent probes such as 6-carboxyfluorescein can be rapidly separated and quantified. However, we also show that only select probes can be used, presumably due to unfavorable interactions between Pol gamma and certain 5' labels.     

基因芯片制备方法研究进展

基因芯片是融微电子学、生命科学和物理学为一体的技术,目前广泛应用于疾病的基因诊断、基因表达研究、基因组研究、发现新基因以及病原体的诊断,具有广阔的应用前景。基因芯片的制备主要可分为原位合成、合成后交联二种方法。本文综述了基因芯片的制备方法分析了各自的优缺点。     

A simple pen-spotting method for arraying biomolecules on solid substrates

We describe a simple, relatively inexpensive method for depositing biomolecules on a solid substrate using Rapidograph® drafting pens. The pens can be used without modification to accurately deposit spots between 100 and 600 μm in diameter. When mounted on a suitable microtranslation stage, the pens can be used to easily deposit tens of spots aligned with underlying substrate features such as microfabricated sensors. The pens are particularly convenient because pre-mixed solutions can be stored in the pens for multiple uses. We demonstrate the use of this approach to deposit DNA probes on a microsensor array.     

Use of DNA fingerprinting for human population genetic studies

DNA fingerprinting techniques have been used in population genetic studies on many different kinds of organisms. Here, we present new applications for multilocus DNA fingerprint probes in population studies and demonstrate the applicability of DNA fingerprinting to human population genetics, using M13 phage DNA as a probe. The new approach, which is based on a factor method of numerical coding of non-quantitative data (factor correspondence analysis-FCA), shows good agreement between population position, as indicated by the three principal factors, and ethnogenetic proximity.     

Immobilisation of DNA probes for the development of SPR-based sensing

An immobilisation procedure based on the direct coupling of thiol-derivatised oligonucleotide probes to bare gold sensor surfaces has been used for DNA sensing applications. The instrumentation used relies on surface plasmon resonance (SPR) transduction; in particular the commercially available instruments BIACORE X and SPREETA, have been employed in this study. The performances of the SPR-based DNA sensors resulting from direct coupling of thiol-derivatised DNA probes onto gold chips, have been studied in terms of the main analytical parameters, i.e. selectivity, sensitivity, reproducibility, analysis time, etc. A comparison between the thiol-derivatised immobilisation approach and a reference immobilisation method, based on the coupling of biotinylated oligonucleotide probes onto a streptavidin coated dextran sensor surface, using synthetic complementary oligonucleotides has been discussed. Finally, a denaturation method to obtain ssDNA ready for hybridisation analysis has been applied to polymerase chain reaction (PCR) amplified samples, for the detection of genetically modified organisms (GMOs).     

SARS病毒再测序DNA微阵列的探针设计

探针设计是SARS病毒再测序DNA微阵列制作的关键步骤,为了保证探针的杂交条件尽可能一致,采用了作者提出的两种等长变覆盖的探针设计方法,即基于Tm距离的算法和遗传算法。针对SAILS病毒基因组中的两段特异序列设计了一组探针,并与等长移位法和变长变覆盖法的设计结果进行了比较。等长变覆盖法得到的探针集在探针长度一致的情况下,探针的Tm值有较小的标准差和变化范围。结果表明,等长变覆盖法得到的探针具有更好的杂交条件一致性。     

Hypervariable single and multi-locus DNA polymorphisms for genetic typing of non-human primates

The series of hypervariable, “minisatellite” loci characterized byJeffreys and coworkers in the human myoglobin gene have proved to be DNA sequences highly conserved throughout the eukaryotic genome, and hence the methodology developed for human DNA “fingerprinting” has found immediate application in an ever expanding number of species. Primatologists have not been slow to profit from a method which predicts individual recognition to a very high degree of probability, and initial studies have focused on paternity allocation (rather than paternity exclusion, as designated by the classical biochemical markers), adaptive aspects of socio-sexual behaviour patterns and mating systems. A number of probes with sequences corresponding to the common minisatellite core sequences have been used for probing genomic DNA, and synthetic, G-rich oligonucleotides (15 – 37 bases), corresponding to the core sequence of the minisatellite repeat unit, or simply di-, tri-, or tetranucleotide repeats, appear to be equally discriminatory. The multiple banding patterns produced on hybridization of these probes to restriction enzyme digests of DNA provide an advantage in that the probability of two unrelated individuals sharing the same banding pattern will be low. However, the uncertainty of linkage of the multiple loci identified precludes genotyping and population genetic analyses based on allele frequencies. In contrast, single locus analysis allows DNA typing using variable number tandem repeat (VNTR) or restriction fragment length (RFLP) DNA polymorphisms, and the merits and drawbacks relative to DNA fingerprinting are discussed. For the behavioural primatologists dealing with defined, accessible troops of primates, the value of multilocus DNA fingerprinting, in terms of established methodology and availability of probes applicable to species as phylogenetically wide-ranging as apes and prosimians, may well outweigh the loss of genotypic and population structure data.     

Existing and emerging detection technologies for DNA (Deoxyribonucleic Acid) finger printing, sequencing, bio- and analytical chips: a multidisciplinary development unifying molecular biology, chemical and electronics engineering

The current status and research trends of detection techniques for DNA-based analysis such as DNA finger printing, sequencing, biochips and allied fields are examined. An overview of main detectors is presented vis-à-vis these DNA operations. The biochip method is explained, the role of micro- and nanoelectronic technologies in biochip realization is highlighted, various optical and electrical detection principles employed in biochips are indicated, and the operational mechanisms of these detection devices are described. Although a diversity of biochips for diagnostic and therapeutic applications has been demonstrated in research laboratories worldwide, only some of these chips have entered the clinical market, and more chips are awaiting commercialization. The necessity of tagging is eliminated in refractive-index change based devices, but the basic flaw of indirect nature of most detection methodologies can only be overcome by generic and/or reagentless DNA sensors such as the conductance-based approach and the DNA-single electron transistor (DNA-SET) structure. Devices of the electrical detection-based category are expected to pave the pathway for the next-generation DNA chips. The review provides a comprehensive coverage of the detection technologies for DNA finger printing, sequencing and related techniques, encompassing a variety of methods from the primitive art to the state-of-the-art scenario as well as promising methods for the future.