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: 7938835
• 负责人: Dmitri Litvinov
• 金额: $ 47.82万
• 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/7938835
• 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)翻译科学和具体的挑战主题，15-RR-101应用翻译技术开发。癌症诊断和预后的挑战呼唤灵敏、特异和经济的探测器系统。特别是基因组学和肿瘤基因组学的快速发展，开启了一个基于DNA和RNA的诊断范围和有效性的巨大扩张期。这些趋势预计将在可预见的未来持续下去，这就要求换能器与杂交分析和分子结合兼容，可采用并行格式实施，对目标分子具有高灵敏度和特异性。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纳克级和微克级样本，而这些样本往往不容易获得。样品放大是一种选择，但可能会导致错误的结果，特别是在不同分析物浓度的比率可能因差异扩增而发生偏差的情况下。迫切需要用于超小临床标本分析的新技术，这将使宝贵的、有限数量的样本得以使用，例如福尔马林固定的石蜡包埋组织、分类的亚群或细针抽吸活检(FNAB)样本。这项拟议工作的目标是开发一种单分子纳米磁性阵列传感器，使此类临床样本能够进行有效的分析。其具体目标是构建一种能够感测单个50 nm磁性标记的纳米磁性传感器阵列，并展示适用于超小型临床标本(如FNAb或分类细胞亚群)的高灵敏度生物分子诊断分析。该设备对低丰度的microRNAs、mRNAs或蛋白质的灵敏度预计将达到前所未有的高水平，可能是在单分子水平上。仅基于一个或几个探针和目标分子进行测量的能力将通过抑制多个相互作用引起的亲和力效应来提高数据质量，并可以揭示目标群体中真正的单分子异质性，而群体平均测量无法检测到。这项研究的基本原理是，这种纳米磁性传感器可以基于快速发展的磁硬盘数据存储技术，并且可以相对容易地集成到具有极高密度的可单独寻址的传感器的实用传感器阵列中。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和蛋白质生物标记物。这项拟议的研究将把生物磁传感推进到一种高度通用的临床诊断技术，它将提供超灵敏的(单分子)分子检测和高特异性，通过磁场拉出熔化来抑制非特异性关联。这项研究有望使新一代高度可靠的分子诊断仪器具有显著增强的灵敏度和高特异度。