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微阵列芯片具有高通量、微量化和自动化等特点,已经在很多领域得到广泛应用。但是微阵列芯片仍然具有不足之处,如所需设备昂贵、分析时间较长、灵敏度不高、多样品平行分析能力不足等。微流控芯片微米级的通道具有相对较大的比表面积和较短的扩散距离,能够显著加快分析速度、提高检测效率、增强分析性能,并且能够加工大量的平行通道用于多样品分析。目前已经有大量文献报道将微流控芯片和微阵列芯片相结合,发展了独特的杂交方式并在实验和理论上分别证明了两者相结合的优势,本文综述了将微流控芯片技术应用于微阵列分析的研究进展,着重介绍了在微流控芯片上进行微阵列分析时的杂交方式、促进杂交的措施以及杂交过程的数学建模,同时也介绍了其他分析步骤方面的进展。最后分析了目前微流控芯片技术在进行微阵列杂交应用方面的不足及其原因,并指出这两项技术相结合的优势和未来。 相似文献
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基于磁珠法核酸提取原理,设计并制作出旋转驱动式核酸提取微流控芯片及自动化平台。微流控芯片包括裂解腔、清洗腔以及洗脱腔等结构,步进电机带动微流控芯片旋转,通过电磁铁吸附微流控芯片内的磁珠,实现磁珠在各腔室转移,完成核酸提取和纯化。对芯片表面疏水性、磁力大小、磁珠分散程度以及核酸洗脱时间进行优化。结果表明,当磁力大小为250 N时,可实现磁珠转移;磁铁放置于芯片上方1 mm时,腔室内磁珠分散程度最好。洗脱时间为20 min时,芯片上提取大肠杆菌的核酸浓度较高。微流控芯片与磁珠核酸提取技术相结合提取的核酸样本,可直接应用于后续聚合酶链式反应扩增环节,有利于实现核酸自动提取及扩增的一体化。 相似文献
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用合适的手段表征生物分子的相互作用对于深刻理解生命过程的本质以及进行医药开发都具有重要意义。将微流控芯片和毛细管电泳相结合的微流控芯片电泳技术具有快速、高效、高通量、样品用量少和易于整合等诸多优势。本文对近年来进行生物分子间相互作用结合常数测定以及结合动力学研究的微流控芯片电泳分离模式、分析方法和芯片检测方法分别做了介绍;简单对比了微流控芯片技术和微阵列生物芯片生物分子间相互作用研究技术;最后分析了微流控芯片技术目前的不足,并对其未来的发展进行了展望。 相似文献
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采用具有紫外光聚合性能的聚乙二醇(PEG)基水凝胶材料, 通过紫外光聚合作用快速加工双层水凝胶微流控芯片, 并验证了其对肿瘤细胞代谢液进行检测的可行性. 与传统微流控芯片材料相比, 该水凝胶芯片材料具有更好的生物相容性及可操控性, 可直接加工成形, 在生物学领域特别是细胞培养过程控制方面具有良好的应用前景. 实验结果表明, 该水凝胶微流控芯片可在微尺度空间有效模拟细胞生长环境, 并实现对细胞连续捕获后的原位培养. 将该芯片与卟啉可视阵列传感器系统结合, 经代谢特征分析可有效区分不同种类肿瘤细胞, 实现芯片细胞培养平台上的细胞代谢指纹快速可视化传感检测. 相似文献
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核酸适配体是短的、单链DNA或RNA序列。相较于抗体成本高、不稳定、免疫原性、难修饰的问题,核酸适配体作为新一代亲和试剂,有着免疫原性低、易修饰、靶标范围广等优势。通过指数富集配体系统进化技术(Systematic Evolution of Ligands by Exponential Enrichment,SELEX)可获得与靶标分子特异性结合的核酸适配体,而如何提高核酸适配体筛选效率是核酸适配体广泛应用的一个瓶颈问题。目前,提高核酸适配体筛选效率的方法种类较多,而微流控SELEX的发展,加速了核酸适配体的发现。核酸适配体作为各种靶标的识别分子,具有诊断和治疗特定疾病的潜力。鉴于此,本文重点介绍核酸适配体在微流控芯片领域进行筛选的研究进展,以及阐述其在病毒性疾病中的应用以及展望。 相似文献
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We describe a rapid, quantitative, multiplex, self-labelled, and real-time DNA biosensor employing Ag nanoparticle-bound DNA hairpin probes immobilized in a microfluidic channel. Capture of complementary target DNAs by the microarrayed DNA hairpin probes results in a positive fluorescence signal via a conformational change of the probe molecules, signalling the presence of target DNAs. The device's capability for quantitative analyses was evaluated and a detection time as low as 6 min (with a target flow rate of 0.5 μl min(-1)) was sufficient to generate significant detection signals. This detection time translates to merely 3 μl of target solution consumption. An unoptimized sensitivity of 500 pM was demonstrated for this device. 相似文献
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Dalavoy TS Wernette DP Gong M Sweedler JV Lu Y Flachsbart BR Shannon MA Bohn PW Cropek DM 《Lab on a chip》2008,8(5):786-793
Due to the numerous toxicological effects of lead, its presence in the environment needs to be effectively monitored. Incorporating a biosensing element within a microfluidic platform enables rapid and reliable determinations of lead at trace levels. A microchip-based lead sensor is described here that employs a lead-specific DNAzyme (also called catalytic DNA or deoxyribozyme) as a recognition element that cleaves its complementary substrate DNA strand only in the presence of cationic lead (Pb(2+)). Fluorescent tags on the DNAzyme translate the cleavage events to measurable, optical signals proportional to Pb(2+) concentration. The DNAzyme responds sensitively and selectively to Pb(2+), and immobilizing DNAzyme in the sensor permits both sensor regeneration and localization of the detection zone. Here, the DNAzyme has been immobilized on a PMMA surface using the highly specific biotin-streptavidin interaction. The strategy includes using streptavidin physisorbed on a PMMA surface to immobilize DNAzyme both on planar PMMA and on the walls of a PMMA microfluidic device. The immobilized DNAzyme retains its Pb(2+) detection activity in the microfluidic device and can be regenerated and reused. The DNAzyme shows no response to other common metal cations and the presence of these contaminants does not interfere with the lead-induced fluorescence signal. While prior work has shown lead-specific catalytic DNA can be used in its solubilized form and while attached to gold substrates to quantitate Pb(2+) in solution, this is the first use of the DNAzyme immobilized within a microfluidic platform for real time Pb(2+) detection. 相似文献
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Recirculating flow accelerates DNA microarray hybridization in a microfluidic device 总被引:1,自引:0,他引:1
A recirculating microfluidic device fabricated by laminating Mylar and glass was developed for the analysis of hybridization of oligonucleotides to DNA microarrays. The device is part of a system that provides controlled hybridization to DNA probes immobilized in a microarray of polyacrylamide gel pads using recirculation and temperature control. The system was used to obtain real-time kinetics of DNA hybridization and more accurate melting profiles of target-probe duplexes than possible using a static hybridization format. Recirculation shortened the time of perfect match target-probe hybridization from 6 hours to 2 hours and shifted the Td by 1.54 degrees C, relative to static conditions. The experimental results were consistent with a three-dimensional simulation of hybridization using a recirculating buffer system. 相似文献
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Microfluidic DNA microarray analysis: a review 总被引:1,自引:0,他引:1
Microarray DNA hybridization techniques have been used widely from basic to applied molecular biology research. Generally, in a DNA microarray, different probe DNA molecules are immobilized on a solid support in groups and form an array of microspots. Then, hybridization to the microarray can be performed by applying sample DNA solutions in either the bulk or the microfluidic manner. Because the immobilized probe DNA binds and retains its complementary target DNA, detection is achieved through the read-out of the tagged markers on the sample target molecules. The recent microfluidic hybridization method shows the advantages of less sample usage and reduced incubation time. Here, sample solutions are confined in microfabricated channels and flow through the probe microarray area. The high surface-to-volume ratio in microchannels of nanolitre volume greatly enhanced the sensitivity as obtained with the bulk solution method. To generate nanolitre flows, different techniques have been developed, and this including electrokinetic control, vacuum suction and syringe pumping. The latter two are pressure-driven methods which are more flexible without the need of considering the physicochemical properties of solutions. Recently, centrifugal force is employed to drive liquid movement in microchannels. This method utilizes the body force from the liquid itself and there are no additional solution interface contacts such as from electrodes or syringes and tubing. Centrifugal force driven flow also features the ease of parallel hybridizations. In this review, we will summarize the recent advances in microfluidic microarray hybridization and compare the applications of various flow methods. 相似文献
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A method for the development of continuous density gradients of immobilized oligonucleotide probes (20mer) along the length of microfluidic channels is demonstrated. The development of continuous density gradients was achieved using variable electrokinetic transport of probes in hybrid glass-polydimethylsiloxane microfluidic chips. The probes were terminated with an amine functional group, and were delivered by electrokinetic pumping to the flat glass channel wall after it had been densely coated with covalently immobilized aldehyde groups. This method provided probe immobilization densities ranging from 4.5(±0.8)×10(13) to 2.5(±0.8)×10(11) molecules cm(-2), with longitudinal dilution and differential mass transport of the injected plug of probes being the primary factors responsible for the gradient of density. The utility of the resulting density gradient of immobilized probes to control the selectivity of hybridization was demonstrated at room temperature by discrimination between a fully complementary oligonucleotide target, and a target strand containing 3 base pair mismatches (3 BPM) based on the spatial pattern of hybridization for sub-picomole quantities of targets. Single nucleotide polymorphism (SNP) discrimination was possible when temperature control was implemented to improve resolution of the mismatch discrimination, allowing SNP discrimination at 35 °C with a contrast ratio of almost 5 to 1. 相似文献
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Sun Y Perch-Nielsen I Dufva M Sabourin D Bang DD Høgberg J Wolff A 《Analytical and bioanalytical chemistry》2012,402(2):741-748
DNA microarrays have become one of the most powerful tools in the field of genomics and medical diagnosis. Recently, there
has been increased interest in combining microfluidics with microarrays since this approach offers advantages in terms of
portability, reduced analysis time, low consumption of reagents, and increased system integration. Polymers are widely used
for microfluidic systems, but fabrication of microarrays on such materials often requires complicated chemical surface modifications,
which hinders the integration of microarrays into microfluidic systems. In this paper, we demonstrate that simple UV irradiation
can be used to directly immobilize poly(T)poly(C)-tagged DNA oligonucleotide probes on many different types of plastics without
any surface modification. On average, five- and fourfold improvement in immobilization and hybridization efficiency have been
achieved compared to surface-modified slides with aminated DNA probes. Moreover, the TC tag only costs 30% of the commonly
used amino group modifications. Using this microarray fabrication technique, a portable cyclic olefin copolymer biochip containing
eight individually addressable microfluidic channels was developed and used for rapid and parallel identification of Avian
Influenza Virus by DNA hybridization. The one-step, cost-effective DNA-linking method on non-modified polymers significantly
simplifies microarray fabrication procedures and permits great flexibility to plastic material selection, thus making it convenient
to integrate microarrays into plastic microfluidic systems. 相似文献
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Hollars CW Puls J Bakajin O Olsan B Talley CE Lane SM Huser T 《Analytical and bioanalytical chemistry》2006,385(8):1384-1388
A rapid bioassay is described based on the detection of colocalized fluorescent DNA probes bound to DNA targets in a pressure-driven
solution flowing through a planar microfluidic channel. By employing total internal reflection excitation of the fluorescent
probes and illumination of almost the entire flow channel, single fluorescent molecules can be efficiently detected leading
to the rapid analysis of nearly the entire solution flowed through the device. Cross-correlation between images obtained from
two spectrally distinct probes is used to determine the target concentration and efficiently reduces the number of false positives.
The rapid analysis of DNA targets in the low pM range in less than a minute is demonstrated. 相似文献
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Portable chemiluminescence multiplex biosensor for quantitative detection of three B19 DNA genotypes
Mara Mirasoli Francesca Bonvicini Luisa Stella Dolci Martina Zangheri Giorgio Gallinella Aldo Roda 《Analytical and bioanalytical chemistry》2013,405(2-3):1139-1143
A miniaturized multiplex biosensor exploiting a microfluidic oligonucleotide array and chemiluminescence (CL) lensless imaging detection has been developed for parvovirus B19 genotyping. The portable device consists of a reaction chip, comprising a glass slide arrayed with three B19 genotype-specific probes and coupled with a polydimethylsiloxane microfluidic layer, and a charge-coupled device camera modified for lensless CL imaging. Immobilized probes were used in DNA hybridization reactions with biotin-labeled targets, and then hybrids were measured by means of an avidin-horseradish peroxidase (HRP) conjugate and CL detection. All hybridization assay procedures have been optimized to be performed at room temperature through the microfluidic elements of the reaction chip, with sample and reagents delivery via capillary force exploiting adsorbent pads to drive fluids along the microchannels. The biosensor enabled multiplex detection of all B19 genotypes, with detectability down to 80 pmol?L?1 for all B19 genotype oligonucleotides and 650 pmol?L?1 for the amplified product of B19 genotype 1, which is comparable with that obtained in traditional PCR-ELISA formats and with notably shorter assay time (30 min vs. 2 h). The specificity of the assay has been evaluated by performing DNA–DNA hybridization reactions among sequences with different degrees of homology, and no cross hybridizations among B19 genotypes have been observed. The clinical applicability has been demonstrated by assaying amplified products obtained from B19 reference serum samples, with results completely consistent with the reference PCR-ELISA method. The next crucial step will be integration in the biosensor of a miniaturized PCR system for DNA amplification and for heat treatment of amplified products. Figure
A portable multiplex biosensor was developed for detection and genotyping of parvovirus B19 DNA, exploiting lensless CL imaging. The reaction chip is composed of a polydimethylsiloxane microfluidic layer coupled with a glass slide on which oligonucleotide probes specific for three different B19 genotypes are covalently immobilized in a 3?×?3 array. The reaction chip was used in hybridization reactions with biotin-labeled targets and then hybrids were then detected by means of an avidin-HRP conjugate, upon addition of a CL substrate for HRP 相似文献
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A Genosensor for Point Mutation Detection of P53 Gene PCR Product Using Magnetic Particles 下载免费PDF全文
Ezat Hamidi‐Asl Jahan Bakhsh Raoof Mohammad Saeid Hejazi Simin Sharifi Seyed Mahdi Golabi Ilaria Palchetti Marco Mascini 《Electroanalysis》2015,27(6):1378-1386
The development of an electrochemical genosensor involving DNA biotinylated capture probe immobilized on streptavidin coated paramagnetic beads and microfluidic based platform for the detection of P53 gene PCR product is reported. The novelty of this work is the combination of a sensitive electrochemical platform and a proper microfluidic system with a simple and effective enzyme signal amplification technology, ELISA, for detection of target DNA sequence and single nucleotide mutation in p53 tumor suppressor gene sequence. The biosensor has been applied to detect the PCR amplified samples and the results shows that it can discriminate successfully perfect matched DNA from mutant form. 相似文献
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We found that DNA probes can be immobilized on anodically prepared porous niobium oxide without a chemical modification of both the DNA probes and the substrate. By using the porous niobium oxide with a positive surface charge, DNA hybridization events are detected on the basis of the blue-shift of a maximum absorption peak in UV-vis-NIR spectroscopy. The blue-shift is ascribed to the change of surface charge upon single- or double-stranded DNA. The method does not require a label and shows high sensitivity with the detection limit of the concentration of 1nM. 相似文献