生物芯片在微生物学研究中的应用

生物芯片是用于检测生物样品中生物分子及生化指标的微小装置,根据其检测靶标不同分为基因芯片、蛋白芯片、细胞芯片、组织芯片及生化芯片;根据其结构和工作原理不同分为阵列型芯片、液体芯片和微流体芯片。由于具有微量化、并行性、快速、高通量和自动化检测的特点,生物芯片被广泛地应用于生物学研究领域。本文重点论述生物芯片在微生物学研究中的应用。     

军事医学科学院生物芯片研究进展

生物芯片技术是20世纪末兴起的以微加工技术、化学和生物技术为依托的一项综合性高新技术,它是目前高新科技水平的代表之一.该技术根据分子间特异性相互作用的原理,将生命科学领域中不连续的分析过程集成于平方厘米大小的固相介质表面或液相介质中构建微分析系统,以实现对蛋白质和核酸等分子的准确、快速、高通量、平行化、自动化及大信息量的检测.基因芯片、蛋白质芯片、组织芯片和芯片实验室都属于生物芯片的范畴.与传统的检测技术相比,生物芯片具有准确、快速和高通量的优势.生物芯片根据其用途可分为实验室研究用和临床体外诊断用两大类.目前,生物芯片技术已广泛应用于疾病分子诊断、药物筛选、食品卫生安全、传染病预防控制、生物反恐和司法鉴定等领域.本文主要从疾病体外诊断的角度对生物芯片技术和产品进行综述.     

常见生物传感芯片及其应用进展

生物传感芯片是一类综合了生物芯片和生物传感器的优点的新型生物芯片,在保持传统生物芯片的高通量、可寻址、并行处理等特点的基础上,与生物传感器技术相结合,进一步提高了芯片检测的灵敏度和特异性。常见的生物传感芯片主要有光纤传感芯片、表面等离子体共振传感芯片、热生物传感芯片、压电晶体传感芯片等,可用于各种生物大分子,如蛋白质、核酸等的检测,金属离子的测定,病原体的检测,药物筛选等。     

生物芯片分类及其技术原理

生物芯片技术针对DNA,RNA,蛋白质及其它生物分子,在芯片上完成一个或多个功能的检测分析。使大量与生命有关的信息集中在一块芯片上,达到对生物分子,细胞,组织的高通量检测分析,本文根据生物芯片的结构与功能特点,将生物芯片分为微点阵（陈列）芯片,微流路芯片两大类,每大类中又进行了较详细地分门别类,同时扼要介绍各自的技术原理,目前生物芯片技术正向着更加微型化和集成化方向发展,芯片实验室代表着更高的发展阶段。     

蛋白质点阵/芯片技术的新进展

蛋白质点阵/芯片技术是分子生物学技术的重要进展,在功能蛋白质组研究方面具有广阔的潜在应用价值.目前发展起来的印迹蛋白微阵列、分子扫描技术和传感器生物芯片质谱,将应用于药靶检测、疾病诊断、蛋白质结构鉴定和/或蛋白质之间的相互作用分析等方面,具有分析速度快、效率高、样品消耗少等特点,将成为生命科学与医学领域新的研究工具.     

生物芯片技术及其在病原微生物检测方面的进展

生物芯片 ( biochip)是二十世纪 90年代中期发展起来的一项尖端技术 [1 ] 。它以玻片、硅为载体 ,在单位面积上高密度地排列大量的生物材料 ,从而达到一次试验同时检测多种疾病或分析多种生物样品的目的。基因芯片、蛋白芯片等都属于生物芯片的范畴。生物芯片根据芯片上的探针不同 ,可分为蛋白芯片和基因芯片。如果芯片上固定的是肽或蛋白 ,则称为肽芯片或蛋白芯片。如果芯片上固定的分子是寡核苷酸探针或靶 DNA,则称为基因芯片。生物芯片是继大规模集成电路之后的又一次具有深远意义的科学技术革命。生物芯片能为现代医学科学及医学诊断…     

生物芯片技术在药物研究中的应用前景

1991年 ,美国Ｓｔｅｐｈｅｎｆｏｄｏｒ等提出了ＤＮＡ芯片的概念[1] ,随着人类基因组计划 (ＨｕｍａｎＧｅｎｏｍｅＰｒｏｊｅｃｔ,ＨＧＰ)的实施 ,生物芯片技术已成为基因组计划中的一种重要技术手段。生物芯片 (ｂｉｏｌｏｇｉｃａｌｃｈｉｐ或ｂｉｏｃｈｉｐ) ,把生化分析系统中的样品制备、生化反应和结果检测三个部分有机地结合起来连续完成。与传统的检测方法相比 ,具有高通量、高信息量、快速、微型化、自动化、成本低、污染少、用途广等特点[2 ] 。生物芯片包括基因芯片、蛋白质芯片或肽芯片、细胞芯片、组织芯片、元件型的微阵列…     

生物芯片技术的原理与应用

生物芯片是指将大量生物讯息密码（寡核苷酸、cDNA、基因组DNA、蛋白质等）以预先设计的方式固定在玻片、硅片等固相载体上组成的密集分子阵列,可分为核酸芯片、蛋白芯片、芯片实验室三类,生物芯片技术的本质是生物信号的平行分析,它利用核酸分子杂交,蛋白分子亲和原理,通过荧光标记技术检测杂交或亲和否,可迅速获得所需信息。高效、快速的生物芯片技术以其无与伦比的优势,在已医学、分子生物学等领域显现出巨大的应用价值,具有非常广阔的发展前景。     

生物芯片国内最新研发进展

我国生物芯片产业初具规模,已经研制成功二百多种生物芯片产品。深圳益生堂研制的丙型肝炎病毒分片段抗体检测试剂（蛋白质片）、北京博奥公司的微阵列芯片扫描仪等六种芯片及设备获得新药证书或医疗器械证书。博奥公司的芯片扫描仪已经出口欧盟、美国,生物芯片和服务销售达二亿元,技术转让和出口近七千万元。中国已经建立了北京国家芯片工程中心、上海国家芯片工程中心、西安微检验工程中心、天津生物芯片公司、南京生物芯片重点实验室共五个生物芯片研发基地。     

生物芯片研究进展

生物芯片是便携式生物化学分析器的核心技术,通过对微加工获得的微束结构作生物化学处理能使成千上万个与生命相关的信息集成在一块厘米见方的芯片上,采用生物芯片可进行生命科学与医学中所主的各种生物化学反应,从而达到对基因、抗原和活体细胞等进行测试分析的目的。生物芯片发展的最终目标是将从样品制备、化学反应到检测的整个生化分析过程集成化以获得所谓的微型全分析系统或称缩微芯片实验室。生物芯片技术的出现将会给     

Photoimmobilization for microarrays

A photoimmobilization method has been developed for the preparation of microarray biochips. This photoimmobilization method makes it possible to easily covalently immobilize various types of organic molecules and cells on a chip. In addition, by using hydrophilic polymers as matrixes, it is possible to reduce nonspecific interactions with biological components. Various proteins, antibodies, and cells have been microarrayed using this technique, and interactions between these proteins, antibodies, and cells have been investigated. This type of microarray biochip will be important for academic applications such as genomics, proteomics, and cellomics, and clinical analyses.     

A flexible light-directed DNA chip synthesis gated by deprotection using solution photogenerated acids

Oligonucleotide microarrays or oDNA chips are effective decoding and analytical tools for genomic sequences and are useful for a broad range of applications. Therefore, it is desirable to have synthesis methods of DNA chips that are highly flexible in sequence design and provide high quality and general adoptability. We report herein, DNA microarray synthesis based on a flexible biochip method. Our method simply uses photogenerated acid (PGA) in solution to trigger deprotection of the 5′-OH group in conventional nucleotide phosphoramidite monomers (i.e. PGA-gated deprotection), with the rest of the reactions in the synthesis cycle the same as those used for routine synthesis of oligonucleotides. The complete DNA chip synthesis process is accomplished on a regular DNA synthesizer that is coupled with a UV-VIS projection display unit for performing digital photolithography. Using this method, oDNA chips containing probes of newly discovered genes can be quickly and easily synthesized at high yields in a conventional laboratory setting. Furthermore, the PGA-gated chemistry should be applicable to microarray syntheses of a variety of combinatorial molecules, such as peptides and organic molecules.     

生物芯片技术

生物芯片技术近年来发展极为迅速。生物芯片这一概念出现在２０世纪８０年代初,９０年代以来随着人类基因组计划研究的深入,生物芯片技术也得以飞速发展。本文将对生物芯片的概念、发展做一全面的叙述,并详细地介绍最新的生物芯片,如ＤＮＡ芯片等的基本原理、分类、制备,以及生物芯片的发展动向和应用前景。     

乳酸菌基因芯片应用研究进展

基因芯片技术是上世纪90年代兴起的一种对成百上千甚至上万个基因同时进行检测的新技术,具有高通量、并行化的特点,广泛应用于基因表达谱测定、基因功能预测、基因突变检测和多态性分析等方面。多种乳酸菌基因组全序列以及其大量EST、16S rDNA、16S-23S基因间区和功能基因序列测定的完成,有力地推动了基因芯片技术在乳酸菌研究中的应用。介绍了基因芯片的基本原理及乳酸菌基因芯片在基因表达、种属鉴定等研究中的应用进展,以期更好地利用和开发乳酸菌基因芯片。     

Development of a protein microarray using sequence-specific DNA binding domain on DNA chip surface

A protein microarray based on DNA microarray platform was developed to identify protein-protein interactions in vitro. The conventional DNA chip surface by 156-bp PCR product was prepared for a substrate of protein microarray. High-affinity sequence-specific DNA binding domain, GAL4 DNA binding domain, was introduced to the protein microarray as fusion partner of a target model protein, enhanced green fluorescent protein. The target protein was oriented immobilized directly on the DNA chip surface. Finally, monoclonal antibody of the target protein was used to identify the immobilized protein on the surface. This study shows that the conventional DNA chip can be used to make a protein microarray directly, and this novel protein microarray can be applicable as a tool for identifying protein-protein interactions.     

Optical Biochip with Multichannels for Detecting Biotin–Streptavidin Based on Localized Surface Plasmon Resonance

A rapid and accurate detection of molecular binding of antigen-antibody signaling in high throughput is of great importance for biosensing technology. We proposed a novel optical biochip with multichannels for the purpose of detection of biotin–streptavidin on the basis of localized surface plasmon resonance. The optical biochip was fabricated using photolithography to form the microarrays functioning with multichannels on glass substrate. There are different nanostructures in each microarray. Dry etching and nanosphere lithography techniques were applied to fabricate Ag nanostructures such as hemispheres, nanocylindricals, triangular, and rhombic nanostructures. We demonstrated that 100-nM target molecule (streptavidin) on these optical biochips can be easily detected by a UV-visible spectrometer. It indicated that period and shape of the nanostructures significantly affect the optical performance of the nanostructures with different shapes and geometrical parameters. Our experimental results demonstrated that the optical biochips with the multichannels can detect the target molecule using the microarrays structured with different shapes and periods simultaneously. Batch processing of immunoassay for different biomolecular through the different channels embedded on the same chip can be realized accordingly.     

Detection of biological macromolecules on a biochip dedicated to UV specific absorption

This work describes an ultraviolet biosensing technique based on specific molecular absorption detected with a previously developed spectrally selective aluminum gallium nitride (AlGaN) based detector. Light absorption signal of DNA and proteins, respectively at 260 nm and 280 nm, is used to image biochips. To allow detection of protein or DNA monolayers at the surface of a biochip, we develop contrast-enhancing multilayer substrates. We analyze them through models and experiments and validate the possibility of measuring absorptions of the order of 10(-3). These multilayer structures display a high reflectivity, and maximize the interaction of the electric field with the biological element at the chip surface. Optimization of the experimental absorption, which includes effects such as roughness of the biochip, spectral and angular resolution of the optics, illumination, etc., is carried out with an inorganic ultraviolet absorber (titanium dioxide) deposit. We obtained an induced absorption contrast enhanced by a factor of 4.0, conferring enough sensitivity to detect monolayers of DNA or proteins. Experimental results on an Escherichia coli histidine-tagged methionyl-tRNA synthetase protein before and after complexation with an anti-polyHis specific antibody validate our biosensing technique. This label-free optical method may be helpful in controlling biochip coatings, and subsequent biological coupling at the surface of a biochip.     

Polymerase chain reaction/ligase detection reaction/hybridization assays using flow-through microfluidic devices for the detection of low-abundant DNA point mutations

We have microfabricated a flow-through biochip for the analysis of single base mutations in genomic DNA using two different materials: (1) a polycarbonate (PC) chip for performing a primary polymerase chain reaction (PCR) followed by an allele-specific ligation detection reaction (LDR) and (2) a poly(methyl methacrylate) (PMMA) chip for the detection of the LDR products using a universal array platform. The operation of the device was demonstrated by detecting low-abundant DNA mutations in gene fragments (K-ras) that carry point mutations with high diagnostic value for colorectal cancers. The PC microchip was used for sequential PCR/LDR in a continuous-flow format, in which the following three steps were carried out: (1) exponential amplification of gene fragments from genomic DNA; (2) mixing of the resultant PCR product with a LDR mixture via a Y-shaped passive micromixer and (3) ligation of two primers only when the particular mutation was present in the genomic DNA. A PMMA chip was employed as the microarray device, where zip code sequences (24-mer), which were complementary to sequences present on the discriminating primer, were micro-printed into fluidic channels embossed into the PMMA substrate. We successfully demonstrate the ability to detect one mutant DNA in 80 normal sequences with the integrated microfluidic device. The PCR/LDR/hybridization assay using the microchips performed the entire assay at a relatively fast processing speed: 18.7 min for PCR, 8.1 min for LDR, 5 min for hybridization, 10 min for washing and 2.6 min for fluorescence scanning (total processing time=ca. 50 min) with an order of magnitude reduction in reagents compared to bench-top formats.     

质粒pCAMBIA1301的检测

用引物延伸芯片法实现对转基因水稻中 生物芯片技术是生物技术和微制造技术的融合, 已广泛用于生命科学的研究及实践、医学科研及临床、药物设计、环境保护、农业、军事等各个领域。而基因芯片是生物芯片中的一种,是指将大量基因探针分子固定于支持物上,然后与标记的样品进行杂交,所以一次可对大量核酸分子进行检测分析,从而解决了传统核酸印迹杂交技术操作复杂、自动化程度低、检测目的分子数量少、效率低等不足。文章探讨了用基因芯片这一新的检测手段对转基因植物的初步检测,采用一种新的反应机制－引物延伸芯片法（arrayed primer extension）,实现了样品扩增和杂交的一步化,而在传统的基因芯片检测中要需要两步来完成,从而为目前基因芯片中大片段样品的检测提供了一种可能性。 Abstract: Biochip technology which had emerged from the fusion of biotechnology and micro/nanofabrication technology at the end of 1980s has been widely used in life science ,medicine,clinical diagnosis,durg design,agricμLture,envioment pretection and strategics. DNA microarray (also call gene chip,DNA chip),one kind of biochips,is small chip containing many oligonucleotide probe .It can hybridize with labelled sample which makes it possible to detect large numbers of oligonucleotides at one time.So DNA microarray can overcome the disadvantage of traditional hybridization technology such as complexity,low automatization,poor efficiency and amount of detcting molecμLes. This paper describes a new method to detect transgenic plant with gene chip.We have developed a novel arrayed-primer extension technique. It combines hybridization and PCR at one step, while two separate steps are needed in the ordinary DNA microarray, therefore our method provide a feasibility to detect long DNA fragment .     

Functionalized self-assembled monolayer on gold for detection of human mitochondrial tRNA gene mutations

We developed a rapid and simple method to identify single-nucleotide polymorphisms (SNPs) in the human mitochondrial tRNA genes. This method is based on a universal, functionalized, self-assembled monolayer, XNA on Gold chip platform. A set of probes sharing a given allele-specific sequence with a single base substitution near the middle of the sequence was immobilized on chips and the chips were then hybridized with fluorescence-labeled reference targets produced by asymmetric polymerase chain reaction from patient DNA. The ratio of the hybridization signals from the reference and test targets with each probe was then calculated. A ratio of above 3 indicates the presence of a wild-type sequence and a ratio of below 0.3 indicates a mutant sequence. We tested the sensitivity of the chip for known mutations in tRNA(Leu(UUR)) and tRNA(Lys) genes and found that it can also be used to discriminate multiple mutations and heteroplasmy, two typical features of human mitochondrial DNA. The XNA on Gold biochip method is a simple and rapid microarray method that can be used to test rapidly and reliably any SNP in the mitochondrial genome or elsewhere. It will be particularly useful for detecting SNPs associated with human diseases.