Single-molecule nanomagnetic assays for ultrasmall sample clinical diagnostics

描述（由申请人提供）：该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敏感、特异和经济的检测系统。基因组学和肿瘤基因组学的快速发展，特别是开启了一个以DNA和rna为基础的诊断的范围和有效性大大扩展的时期。这些趋势预计将在可预见的未来持续下去，需要与杂交分析和分子结合兼容的换能器，可以并行格式实现，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样品，在纳克和微克水平上，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，特别是当不同分析物浓度的比例可能被差分扩增所扭曲时。目前迫切需要超小型临床标本分析的新技术，这将能够使用珍贵的，有限数量的样本，如福尔马林固定石蜡包埋组织，分类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。具体目标是建立一个纳米磁传感器阵列，能够感应单个50nm磁标签，并演示适用于超小型临床标本（如FNAB或分选细胞亚群）的高灵敏度生物分子诊断分析。该设备对低丰度microRNAs、mrna或蛋白质的灵敏度有望达到前所未有的高，可能达到单分子水平。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由多重相互作用引起的贪婪效应来提高数据质量，并且可以揭示目标群体中无法通过群体平均测量检测到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单个可寻址传感器的实用传感器阵列中。这项研究的影响是，它将使新一代的生物传感器能够高度可靠和敏感地检测和表征mRNA， miRNA和蛋白质生物标志物。提出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，该技术将提供超灵敏（单分子）分子检测和高特异性，通过磁场拉脱熔化来抑制非特异性关联。这项研究有望使新一代高可靠的分子诊断仪器具有显著提高的灵敏度和高特异性。

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.