Single-molecule nanomagnetic assays for ultrasmall sample clinical diagnostics

用于超小样本临床诊断的单分子纳米磁性测定

• 批准号: 7827470
• 负责人: Dmitri Litvinov
• 金额: $ 48.37万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-24 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7827470
• 关键词: Address; Affinity; Antibody Affinity; Archives; Area; Avidity; Binding; Biological Assay; Biological Markers; Biopsy; Biopsy Specimen; Biosensor; Cancer Prognosis; Cell Separation; Cells; Clinical; Complex; DNA; DNA Probes; Data; Data Quality; Data Storage and Retrieval; Detection; Device Designs; Devices; Diagnostic; Effectiveness; Fine needle aspiration biopsy; Flushing; Formalin; Future; Generations; Genomics; Goals; Growth; Heterogeneity; Human; Individual; Label; Lead; Legal patent; Magnetism; Malignant Neoplasms; Measurement; Medical; Messenger RNA; Methods; MicroRNAs; Miniaturization; Modeling; Molecular; Outcome; Paraffin Embedding; Population; Proteins; Proteomics; Reading; Research; Resolution; Sampling; Sensitivity and Specificity; Signal Transduction; Sorting - Cell Movement; Specificity; Specimen; Surface; System; Target Populations; Technology; Testing; Tissues; Transducers; Translational Research; Translations; United States National Institutes of Health; Universities; Work; base; cancer diagnosis; cost; density; design; detector; experience; genome-wide; improved; innovation; instrumentation; magnetic field; melting; member; molecular recognition; nanolabel; nanomagnetic; nanoparticle; nanoscale; new technology; outcome forecast; particle; professor; sensor; single molecule; submicron; technology development; trend

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (15) Translational Science and specific Challenge Topic, 15-RR-101 Applied Translational Technology Development. The challenges of cancer diagnosis and prognosis call for sensitive, specific, and economical detector systems. Rapid advances in genomics and oncogenomics, in particular, are opening a period of great expansion in the range and effectiveness of DNA- and RNA-based diagnostics. These trends, which are expected to continue for the foreseeable future, call for transducers compatible with hybridization assays and molecular binding, implementable in parallel formats, with high sensitivity and specificity for target molecules. While current biomolecular recognition technologies can carry out genome-wide profiling of clinical specimens, they require relatively large samples, at the nanogram and microgram levels, which are often not readily available. Sample amplification is an option but can lead to erroneous results, especially where ratios of different analytes' concentrations can be skewed by differential amplification. There exists a critical need for novel technologies for ultra-small clinical specimen analysis, which will enable the use of precious, limited-amount samples such as formalin-fixed paraffin embedded tissues, sorted sub-populations or fine needle aspirate biopsy (FNAB) samples. The goal of the proposed work is to develop a single-molecule nanomagnetic array sensor that will enable efficient analysis of such clinical samples. The specific objective is to build a nanomagnetic sensor array capable of sensing single 50nm magnetic labels and to demonstrate high-sensitivity biomolecular diagnostic assays applicable to ultra-small clinical specimens such as FNAB or sorted cell sub-populations. The sensitivity of the device to low-abundance microRNAs, mRNAs or proteins is expected to be unprecedentedly high, potentially at the single-molecule level. The ability to base measurements on only one or a few probe and target molecules will improve the quality of the data by suppressing avidity effects arising from multiple interactions, and can reveal genuine single-molecule heterogeneity in target populations not detectable by population-averaged measurements. The rationale for this research is that such a nanomagnetic sensor can be based on rapidly-advancing magnetic hard disk data storage technology, and can be relatively easily integrated into a practical sensor array with an extremely high density of individually-addressable sensors. The impact of this research is that it will enable a new generation of biosensors capable of highly reliable and sensitive detection and characterization of mRNA, miRNA and protein biomarkers. The proposed research will advance biomagnetic sensing into a highly versatile clinical diagnostic technology, which will offer ultrasensitive (single-molecule) molecular detection and high specificity via magnetic field pull off melting to suppress non-specific associations. This research is expected to enable a new generation of highly reliable molecular diagnostic instrumentation with significantly enhanced sensitivity and high specificity.

描述（由申请人提供）：本申请涉及广泛的挑战领域（15）转化科学和特定的挑战主题，15-RR-101应用转化技术开发。癌症诊断和预后的挑战需要灵敏、特异和经济的检测系统。特别是基因组学和癌基因组学的快速发展，正在开启一个基于DNA和RNA的诊断的范围和有效性大幅扩展的时期。这些趋势，预计将继续在可预见的未来，要求与杂交测定和分子结合兼容的转换器，可实施的并行格式，具有高灵敏度和特异性的目标分子。虽然目前的生物分子识别技术可以对临床标本进行全基因组分析，但它们需要相对较大的样品，在纳克和微克水平，这通常是不容易获得的。样品扩增是一种选择，但可能导致错误的结果，特别是在不同分析物浓度的比率可能因差异扩增而偏斜的情况下。存在对用于超小临床样本分析的新技术的迫切需求，这将使得能够使用珍贵的、有限量的样本，诸如福尔马林固定的石蜡包埋的组织、分选的亚群或细针抽吸活检（FNAB）样本。这项工作的目标是开发一种单分子纳米磁性阵列传感器，使这种临床样品的有效分析成为可能。具体目标是构建能够感测单个50 nm磁性标签的纳米磁性传感器阵列，并展示适用于超小临床标本（如FNAB或分选的细胞亚群）的高灵敏度生物分子诊断测定。该设备对低丰度microRNA、mRNA或蛋白质的灵敏度预计将前所未有地高，可能在单分子水平上。基于仅一个或几个探针和靶分子的测量的能力将通过抑制由多种相互作用引起的亲合力效应来提高数据的质量，并且可以揭示通过群体平均测量无法检测到的靶群体中的真正的单分子异质性。这项研究的基本原理是，这种纳米磁传感器可以基于快速发展的磁性硬盘数据存储技术，并且可以相对容易地集成到具有极高密度的可单独寻址传感器的实际传感器阵列中。这项研究的影响是，它将使新一代生物传感器能够高度可靠和灵敏地检测和表征mRNA，miRNA和蛋白质生物标志物。拟议的研究将推动生物磁传感成为一种高度通用的临床诊断技术，该技术将通过磁场拉脱熔化提供超灵敏（单分子）分子检测和高特异性，以抑制非特异性关联。这项研究有望使新一代高度可靠的分子诊断仪器具有显着增强的灵敏度和高特异性。